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+anno_start	anno_end	anno_text	entity_type	sentence	section
+41	46	Cdc42	protein	Investigation of the Interaction between Cdc42 and Its Effector TOCA1	TITLE
+64	69	TOCA1	protein	Investigation of the Interaction between Cdc42 and Its Effector TOCA1	TITLE
+0	54	Transducer of Cdc42-dependent actin assembly protein 1	protein	Transducer of Cdc42-dependent actin assembly protein 1 (TOCA1) is an effector of the Rho family small G protein Cdc42.	ABSTRACT
+56	61	TOCA1	protein	Transducer of Cdc42-dependent actin assembly protein 1 (TOCA1) is an effector of the Rho family small G protein Cdc42.	ABSTRACT
+85	111	Rho family small G protein	protein_type	Transducer of Cdc42-dependent actin assembly protein 1 (TOCA1) is an effector of the Rho family small G protein Cdc42.	ABSTRACT
+112	117	Cdc42	protein	Transducer of Cdc42-dependent actin assembly protein 1 (TOCA1) is an effector of the Rho family small G protein Cdc42.	ABSTRACT
+33	38	F-BAR	structure_element	It contains a membrane-deforming F-BAR domain as well as a Src homology 3 (SH3) domain and a G protein-binding homology region 1 (HR1) domain.	ABSTRACT
+59	73	Src homology 3	structure_element	It contains a membrane-deforming F-BAR domain as well as a Src homology 3 (SH3) domain and a G protein-binding homology region 1 (HR1) domain.	ABSTRACT
+75	78	SH3	structure_element	It contains a membrane-deforming F-BAR domain as well as a Src homology 3 (SH3) domain and a G protein-binding homology region 1 (HR1) domain.	ABSTRACT
+93	128	G protein-binding homology region 1	structure_element	It contains a membrane-deforming F-BAR domain as well as a Src homology 3 (SH3) domain and a G protein-binding homology region 1 (HR1) domain.	ABSTRACT
+130	133	HR1	structure_element	It contains a membrane-deforming F-BAR domain as well as a Src homology 3 (SH3) domain and a G protein-binding homology region 1 (HR1) domain.	ABSTRACT
+0	5	TOCA1	protein	TOCA1 binding to Cdc42 leads to actin rearrangements, which are thought to be involved in processes such as endocytosis, filopodia formation, and cell migration.	ABSTRACT
+17	22	Cdc42	protein	TOCA1 binding to Cdc42 leads to actin rearrangements, which are thought to be involved in processes such as endocytosis, filopodia formation, and cell migration.	ABSTRACT
+8	14	solved	experimental_method	We have solved the structure of the HR1 domain of TOCA1, providing the first structural data for this protein.	ABSTRACT
+19	28	structure	evidence	We have solved the structure of the HR1 domain of TOCA1, providing the first structural data for this protein.	ABSTRACT
+36	39	HR1	structure_element	We have solved the structure of the HR1 domain of TOCA1, providing the first structural data for this protein.	ABSTRACT
+50	55	TOCA1	protein	We have solved the structure of the HR1 domain of TOCA1, providing the first structural data for this protein.	ABSTRACT
+77	92	structural data	evidence	We have solved the structure of the HR1 domain of TOCA1, providing the first structural data for this protein.	ABSTRACT
+23	28	TOCA1	protein	We have found that the TOCA1 HR1, like the closely related CIP4 HR1, has interesting structural features that are not observed in other HR1 domains.	ABSTRACT
+29	32	HR1	structure_element	We have found that the TOCA1 HR1, like the closely related CIP4 HR1, has interesting structural features that are not observed in other HR1 domains.	ABSTRACT
+59	63	CIP4	protein	We have found that the TOCA1 HR1, like the closely related CIP4 HR1, has interesting structural features that are not observed in other HR1 domains.	ABSTRACT
+64	67	HR1	structure_element	We have found that the TOCA1 HR1, like the closely related CIP4 HR1, has interesting structural features that are not observed in other HR1 domains.	ABSTRACT
+136	139	HR1	structure_element	We have found that the TOCA1 HR1, like the closely related CIP4 HR1, has interesting structural features that are not observed in other HR1 domains.	ABSTRACT
+45	49	TOCA	protein	We have also investigated the binding of the TOCA HR1 domain to Cdc42 and the potential ternary complex between Cdc42 and the G protein-binding regions of TOCA1 and a member of the Wiskott-Aldrich syndrome protein family, N-WASP.	ABSTRACT
+50	53	HR1	structure_element	We have also investigated the binding of the TOCA HR1 domain to Cdc42 and the potential ternary complex between Cdc42 and the G protein-binding regions of TOCA1 and a member of the Wiskott-Aldrich syndrome protein family, N-WASP.	ABSTRACT
+64	69	Cdc42	protein	We have also investigated the binding of the TOCA HR1 domain to Cdc42 and the potential ternary complex between Cdc42 and the G protein-binding regions of TOCA1 and a member of the Wiskott-Aldrich syndrome protein family, N-WASP.	ABSTRACT
+112	117	Cdc42	protein	We have also investigated the binding of the TOCA HR1 domain to Cdc42 and the potential ternary complex between Cdc42 and the G protein-binding regions of TOCA1 and a member of the Wiskott-Aldrich syndrome protein family, N-WASP.	ABSTRACT
+126	151	G protein-binding regions	site	We have also investigated the binding of the TOCA HR1 domain to Cdc42 and the potential ternary complex between Cdc42 and the G protein-binding regions of TOCA1 and a member of the Wiskott-Aldrich syndrome protein family, N-WASP.	ABSTRACT
+155	160	TOCA1	protein	We have also investigated the binding of the TOCA HR1 domain to Cdc42 and the potential ternary complex between Cdc42 and the G protein-binding regions of TOCA1 and a member of the Wiskott-Aldrich syndrome protein family, N-WASP.	ABSTRACT
+181	220	Wiskott-Aldrich syndrome protein family	protein_type	We have also investigated the binding of the TOCA HR1 domain to Cdc42 and the potential ternary complex between Cdc42 and the G protein-binding regions of TOCA1 and a member of the Wiskott-Aldrich syndrome protein family, N-WASP.	ABSTRACT
+222	228	N-WASP	protein	We have also investigated the binding of the TOCA HR1 domain to Cdc42 and the potential ternary complex between Cdc42 and the G protein-binding regions of TOCA1 and a member of the Wiskott-Aldrich syndrome protein family, N-WASP.	ABSTRACT
+0	5	TOCA1	protein	TOCA1 binds Cdc42 with micromolar affinity, in contrast to the nanomolar affinity of the N-WASP G protein-binding region for Cdc42.	ABSTRACT
+12	17	Cdc42	protein	TOCA1 binds Cdc42 with micromolar affinity, in contrast to the nanomolar affinity of the N-WASP G protein-binding region for Cdc42.	ABSTRACT
+89	95	N-WASP	protein	TOCA1 binds Cdc42 with micromolar affinity, in contrast to the nanomolar affinity of the N-WASP G protein-binding region for Cdc42.	ABSTRACT
+96	120	G protein-binding region	site	TOCA1 binds Cdc42 with micromolar affinity, in contrast to the nanomolar affinity of the N-WASP G protein-binding region for Cdc42.	ABSTRACT
+125	130	Cdc42	protein	TOCA1 binds Cdc42 with micromolar affinity, in contrast to the nanomolar affinity of the N-WASP G protein-binding region for Cdc42.	ABSTRACT
+0	3	NMR	experimental_method	NMR experiments show that the Cdc42-binding domain from N-WASP is able to displace TOCA1 HR1 from Cdc42, whereas the N-WASP domain but not the TOCA1 HR1 domain inhibits actin polymerization.	ABSTRACT
+30	50	Cdc42-binding domain	site	NMR experiments show that the Cdc42-binding domain from N-WASP is able to displace TOCA1 HR1 from Cdc42, whereas the N-WASP domain but not the TOCA1 HR1 domain inhibits actin polymerization.	ABSTRACT
+56	62	N-WASP	protein	NMR experiments show that the Cdc42-binding domain from N-WASP is able to displace TOCA1 HR1 from Cdc42, whereas the N-WASP domain but not the TOCA1 HR1 domain inhibits actin polymerization.	ABSTRACT
+83	88	TOCA1	protein	NMR experiments show that the Cdc42-binding domain from N-WASP is able to displace TOCA1 HR1 from Cdc42, whereas the N-WASP domain but not the TOCA1 HR1 domain inhibits actin polymerization.	ABSTRACT
+89	92	HR1	structure_element	NMR experiments show that the Cdc42-binding domain from N-WASP is able to displace TOCA1 HR1 from Cdc42, whereas the N-WASP domain but not the TOCA1 HR1 domain inhibits actin polymerization.	ABSTRACT
+98	103	Cdc42	protein	NMR experiments show that the Cdc42-binding domain from N-WASP is able to displace TOCA1 HR1 from Cdc42, whereas the N-WASP domain but not the TOCA1 HR1 domain inhibits actin polymerization.	ABSTRACT
+117	123	N-WASP	protein	NMR experiments show that the Cdc42-binding domain from N-WASP is able to displace TOCA1 HR1 from Cdc42, whereas the N-WASP domain but not the TOCA1 HR1 domain inhibits actin polymerization.	ABSTRACT
+143	148	TOCA1	protein	NMR experiments show that the Cdc42-binding domain from N-WASP is able to displace TOCA1 HR1 from Cdc42, whereas the N-WASP domain but not the TOCA1 HR1 domain inhibits actin polymerization.	ABSTRACT
+149	152	HR1	structure_element	NMR experiments show that the Cdc42-binding domain from N-WASP is able to displace TOCA1 HR1 from Cdc42, whereas the N-WASP domain but not the TOCA1 HR1 domain inhibits actin polymerization.	ABSTRACT
+19	24	TOCA1	protein	This suggests that TOCA1 binding to Cdc42 is an early step in the Cdc42-dependent pathways that govern actin dynamics, and the differential binding affinities of the effectors facilitate a handover from TOCA1 to N-WASP, which can then drive recruitment of the actin-modifying machinery.	ABSTRACT
+36	41	Cdc42	protein	This suggests that TOCA1 binding to Cdc42 is an early step in the Cdc42-dependent pathways that govern actin dynamics, and the differential binding affinities of the effectors facilitate a handover from TOCA1 to N-WASP, which can then drive recruitment of the actin-modifying machinery.	ABSTRACT
+66	71	Cdc42	protein	This suggests that TOCA1 binding to Cdc42 is an early step in the Cdc42-dependent pathways that govern actin dynamics, and the differential binding affinities of the effectors facilitate a handover from TOCA1 to N-WASP, which can then drive recruitment of the actin-modifying machinery.	ABSTRACT
+140	158	binding affinities	evidence	This suggests that TOCA1 binding to Cdc42 is an early step in the Cdc42-dependent pathways that govern actin dynamics, and the differential binding affinities of the effectors facilitate a handover from TOCA1 to N-WASP, which can then drive recruitment of the actin-modifying machinery.	ABSTRACT
+203	208	TOCA1	protein	This suggests that TOCA1 binding to Cdc42 is an early step in the Cdc42-dependent pathways that govern actin dynamics, and the differential binding affinities of the effectors facilitate a handover from TOCA1 to N-WASP, which can then drive recruitment of the actin-modifying machinery.	ABSTRACT
+212	218	N-WASP	protein	This suggests that TOCA1 binding to Cdc42 is an early step in the Cdc42-dependent pathways that govern actin dynamics, and the differential binding affinities of the effectors facilitate a handover from TOCA1 to N-WASP, which can then drive recruitment of the actin-modifying machinery.	ABSTRACT
+4	19	Ras superfamily	protein_type	The Ras superfamily of small GTPases comprises over 150 members that regulate a multitude of cellular processes in eukaryotes.	INTRO
+23	36	small GTPases	protein_type	The Ras superfamily of small GTPases comprises over 150 members that regulate a multitude of cellular processes in eukaryotes.	INTRO
+115	125	eukaryotes	taxonomy_domain	The Ras superfamily of small GTPases comprises over 150 members that regulate a multitude of cellular processes in eukaryotes.	INTRO
+99	102	Ras	protein_type	The superfamily can be divided into five families based on structural and functional similarities: Ras, Rho, Rab, Arf, and Ran.	INTRO
+104	107	Rho	protein_type	The superfamily can be divided into five families based on structural and functional similarities: Ras, Rho, Rab, Arf, and Ran.	INTRO
+109	112	Rab	protein_type	The superfamily can be divided into five families based on structural and functional similarities: Ras, Rho, Rab, Arf, and Ran.	INTRO
+114	117	Arf	protein_type	The superfamily can be divided into five families based on structural and functional similarities: Ras, Rho, Rab, Arf, and Ran.	INTRO
+123	126	Ran	protein_type	The superfamily can be divided into five families based on structural and functional similarities: Ras, Rho, Rab, Arf, and Ran.	INTRO
+72	80	G domain	structure_element	All members share a well defined core structure of ∼20 kDa known as the G domain, which is responsible for guanine nucleotide binding.	INTRO
+107	125	guanine nucleotide	chemical	All members share a well defined core structure of ∼20 kDa known as the G domain, which is responsible for guanine nucleotide binding.	INTRO
+39	45	active	protein_state	These molecular switches cycle between active, GTP-bound, and inactive, GDP-bound, states with the help of auxiliary proteins.	INTRO
+47	56	GTP-bound	protein_state	These molecular switches cycle between active, GTP-bound, and inactive, GDP-bound, states with the help of auxiliary proteins.	INTRO
+62	70	inactive	protein_state	These molecular switches cycle between active, GTP-bound, and inactive, GDP-bound, states with the help of auxiliary proteins.	INTRO
+72	81	GDP-bound	protein_state	These molecular switches cycle between active, GTP-bound, and inactive, GDP-bound, states with the help of auxiliary proteins.	INTRO
+4	39	guanine nucleotide exchange factors	protein_type	The guanine nucleotide exchange factors mediate formation of the active state by promoting the dissociation of GDP, allowing GTP to bind.	INTRO
+65	71	active	protein_state	The guanine nucleotide exchange factors mediate formation of the active state by promoting the dissociation of GDP, allowing GTP to bind.	INTRO
+111	114	GDP	chemical	The guanine nucleotide exchange factors mediate formation of the active state by promoting the dissociation of GDP, allowing GTP to bind.	INTRO
+125	128	GTP	chemical	The guanine nucleotide exchange factors mediate formation of the active state by promoting the dissociation of GDP, allowing GTP to bind.	INTRO
+4	30	GTPase-activating proteins	protein_type	The GTPase-activating proteins stimulate the rate of intrinsic GTP hydrolysis, mediating the return to the inactive state (reviewed in Ref.).	INTRO
+63	66	GTP	chemical	The GTPase-activating proteins stimulate the rate of intrinsic GTP hydrolysis, mediating the return to the inactive state (reviewed in Ref.).	INTRO
+107	115	inactive	protein_state	The GTPase-activating proteins stimulate the rate of intrinsic GTP hydrolysis, mediating the return to the inactive state (reviewed in Ref.).	INTRO
+28	44	small G proteins	protein_type	The overall conformation of small G proteins in the active and inactive states is similar, but they differ significantly in two main regions known as switch I and switch II.	INTRO
+52	58	active	protein_state	The overall conformation of small G proteins in the active and inactive states is similar, but they differ significantly in two main regions known as switch I and switch II.	INTRO
+63	71	inactive	protein_state	The overall conformation of small G proteins in the active and inactive states is similar, but they differ significantly in two main regions known as switch I and switch II.	INTRO
+150	158	switch I	site	The overall conformation of small G proteins in the active and inactive states is similar, but they differ significantly in two main regions known as switch I and switch II.	INTRO
+163	172	switch II	site	The overall conformation of small G proteins in the active and inactive states is similar, but they differ significantly in two main regions known as switch I and switch II.	INTRO
+75	84	GTP-bound	protein_state	These regions are responsible for “sensing” the nucleotide state, with the GTP-bound state showing greater rigidity and the GDP-bound state adopting a more relaxed conformation (reviewed in Ref.).	INTRO
+124	133	GDP-bound	protein_state	These regions are responsible for “sensing” the nucleotide state, with the GTP-bound state showing greater rigidity and the GDP-bound state adopting a more relaxed conformation (reviewed in Ref.).	INTRO
+7	13	active	protein_state	In the active state, G proteins bind to an array of downstream effectors, through which they exert their extensive roles within the cell.	INTRO
+21	31	G proteins	protein_type	In the active state, G proteins bind to an array of downstream effectors, through which they exert their extensive roles within the cell.	INTRO
+4	14	structures	evidence	The structures of more than 60 small G protein·effector complexes have been solved, and, not surprisingly, the switch regions have been implicated in a large proportion of the G protein-effector interactions (reviewed in Ref.).	INTRO
+37	46	G protein	protein_type	The structures of more than 60 small G protein·effector complexes have been solved, and, not surprisingly, the switch regions have been implicated in a large proportion of the G protein-effector interactions (reviewed in Ref.).	INTRO
+76	82	solved	experimental_method	The structures of more than 60 small G protein·effector complexes have been solved, and, not surprisingly, the switch regions have been implicated in a large proportion of the G protein-effector interactions (reviewed in Ref.).	INTRO
+111	125	switch regions	site	The structures of more than 60 small G protein·effector complexes have been solved, and, not surprisingly, the switch regions have been implicated in a large proportion of the G protein-effector interactions (reviewed in Ref.).	INTRO
+176	185	G protein	protein_type	The structures of more than 60 small G protein·effector complexes have been solved, and, not surprisingly, the switch regions have been implicated in a large proportion of the G protein-effector interactions (reviewed in Ref.).	INTRO
+134	143	G protein	protein_type	However, because each of the 150 members of the superfamily interacts with multiple effectors, there are still a huge number of known G protein-effector interactions that have not yet been studied structurally.	INTRO
+4	14	Rho family	protein_type	The Rho family comprises 20 members, of which three, RhoA, Rac1, and Cdc42, have been relatively well studied.	INTRO
+53	57	RhoA	protein	The Rho family comprises 20 members, of which three, RhoA, Rac1, and Cdc42, have been relatively well studied.	INTRO
+59	63	Rac1	protein	The Rho family comprises 20 members, of which three, RhoA, Rac1, and Cdc42, have been relatively well studied.	INTRO
+69	74	Cdc42	protein	The Rho family comprises 20 members, of which three, RhoA, Rac1, and Cdc42, have been relatively well studied.	INTRO
+0	4	RhoA	protein	RhoA acts to rearrange existing actin structures to form stress fibers, whereas Rac1 and Cdc42 promote de novo actin polymerization to form lamellipodia and filopodia, respectively.	INTRO
+80	84	Rac1	protein	RhoA acts to rearrange existing actin structures to form stress fibers, whereas Rac1 and Cdc42 promote de novo actin polymerization to form lamellipodia and filopodia, respectively.	INTRO
+89	94	Cdc42	protein	RhoA acts to rearrange existing actin structures to form stress fibers, whereas Rac1 and Cdc42 promote de novo actin polymerization to form lamellipodia and filopodia, respectively.	INTRO
+12	16	RhoA	protein	A number of RhoA and Rac1 effector proteins, including the formins and members of the protein kinase C-related kinase (PRK)6 family, along with Cdc42 effectors, including the Wiskott-Aldrich syndrome (WASP) family and the transducer of Cdc42-dependent actin assembly (TOCA) family, have also been linked to the pathways that govern cytoskeletal dynamics.	INTRO
+21	25	Rac1	protein	A number of RhoA and Rac1 effector proteins, including the formins and members of the protein kinase C-related kinase (PRK)6 family, along with Cdc42 effectors, including the Wiskott-Aldrich syndrome (WASP) family and the transducer of Cdc42-dependent actin assembly (TOCA) family, have also been linked to the pathways that govern cytoskeletal dynamics.	INTRO
+86	117	protein kinase C-related kinase	protein_type	A number of RhoA and Rac1 effector proteins, including the formins and members of the protein kinase C-related kinase (PRK)6 family, along with Cdc42 effectors, including the Wiskott-Aldrich syndrome (WASP) family and the transducer of Cdc42-dependent actin assembly (TOCA) family, have also been linked to the pathways that govern cytoskeletal dynamics.	INTRO
+119	122	PRK	protein_type	A number of RhoA and Rac1 effector proteins, including the formins and members of the protein kinase C-related kinase (PRK)6 family, along with Cdc42 effectors, including the Wiskott-Aldrich syndrome (WASP) family and the transducer of Cdc42-dependent actin assembly (TOCA) family, have also been linked to the pathways that govern cytoskeletal dynamics.	INTRO
+123	124	6	protein_type	A number of RhoA and Rac1 effector proteins, including the formins and members of the protein kinase C-related kinase (PRK)6 family, along with Cdc42 effectors, including the Wiskott-Aldrich syndrome (WASP) family and the transducer of Cdc42-dependent actin assembly (TOCA) family, have also been linked to the pathways that govern cytoskeletal dynamics.	INTRO
+144	149	Cdc42	protein	A number of RhoA and Rac1 effector proteins, including the formins and members of the protein kinase C-related kinase (PRK)6 family, along with Cdc42 effectors, including the Wiskott-Aldrich syndrome (WASP) family and the transducer of Cdc42-dependent actin assembly (TOCA) family, have also been linked to the pathways that govern cytoskeletal dynamics.	INTRO
+175	199	Wiskott-Aldrich syndrome	protein_type	A number of RhoA and Rac1 effector proteins, including the formins and members of the protein kinase C-related kinase (PRK)6 family, along with Cdc42 effectors, including the Wiskott-Aldrich syndrome (WASP) family and the transducer of Cdc42-dependent actin assembly (TOCA) family, have also been linked to the pathways that govern cytoskeletal dynamics.	INTRO
+201	205	WASP	protein_type	A number of RhoA and Rac1 effector proteins, including the formins and members of the protein kinase C-related kinase (PRK)6 family, along with Cdc42 effectors, including the Wiskott-Aldrich syndrome (WASP) family and the transducer of Cdc42-dependent actin assembly (TOCA) family, have also been linked to the pathways that govern cytoskeletal dynamics.	INTRO
+236	266	Cdc42-dependent actin assembly	protein_type	A number of RhoA and Rac1 effector proteins, including the formins and members of the protein kinase C-related kinase (PRK)6 family, along with Cdc42 effectors, including the Wiskott-Aldrich syndrome (WASP) family and the transducer of Cdc42-dependent actin assembly (TOCA) family, have also been linked to the pathways that govern cytoskeletal dynamics.	INTRO
+268	272	TOCA	protein_type	A number of RhoA and Rac1 effector proteins, including the formins and members of the protein kinase C-related kinase (PRK)6 family, along with Cdc42 effectors, including the Wiskott-Aldrich syndrome (WASP) family and the transducer of Cdc42-dependent actin assembly (TOCA) family, have also been linked to the pathways that govern cytoskeletal dynamics.	INTRO
+0	5	Cdc42	protein	Cdc42 effectors, TOCA1 and the ubiquitously expressed member of the WASP family, N-WASP, have been implicated in the regulation of actin polymerization downstream of Cdc42 and phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2).	INTRO
+17	22	TOCA1	protein	Cdc42 effectors, TOCA1 and the ubiquitously expressed member of the WASP family, N-WASP, have been implicated in the regulation of actin polymerization downstream of Cdc42 and phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2).	INTRO
+68	79	WASP family	protein_type	Cdc42 effectors, TOCA1 and the ubiquitously expressed member of the WASP family, N-WASP, have been implicated in the regulation of actin polymerization downstream of Cdc42 and phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2).	INTRO
+81	87	N-WASP	protein	Cdc42 effectors, TOCA1 and the ubiquitously expressed member of the WASP family, N-WASP, have been implicated in the regulation of actin polymerization downstream of Cdc42 and phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2).	INTRO
+166	171	Cdc42	protein	Cdc42 effectors, TOCA1 and the ubiquitously expressed member of the WASP family, N-WASP, have been implicated in the regulation of actin polymerization downstream of Cdc42 and phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2).	INTRO
+176	213	phosphatidylinositol 4,5-bisphosphate	chemical	Cdc42 effectors, TOCA1 and the ubiquitously expressed member of the WASP family, N-WASP, have been implicated in the regulation of actin polymerization downstream of Cdc42 and phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2).	INTRO
+215	224	PI(4,5)P2	chemical	Cdc42 effectors, TOCA1 and the ubiquitously expressed member of the WASP family, N-WASP, have been implicated in the regulation of actin polymerization downstream of Cdc42 and phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2).	INTRO
+0	6	N-WASP	protein	N-WASP exists in an autoinhibited conformation, which is released upon PI(4,5)P2 and Cdc42 binding or by other factors, such as phosphorylation.	INTRO
+20	46	autoinhibited conformation	protein_state	N-WASP exists in an autoinhibited conformation, which is released upon PI(4,5)P2 and Cdc42 binding or by other factors, such as phosphorylation.	INTRO
+71	80	PI(4,5)P2	chemical	N-WASP exists in an autoinhibited conformation, which is released upon PI(4,5)P2 and Cdc42 binding or by other factors, such as phosphorylation.	INTRO
+85	90	Cdc42	protein	N-WASP exists in an autoinhibited conformation, which is released upon PI(4,5)P2 and Cdc42 binding or by other factors, such as phosphorylation.	INTRO
+29	47	C-terminal regions	structure_element	Following their release, the C-terminal regions of N-WASP are free to interact with G-actin and a known nucleator of actin assembly, the Arp2/3 complex.	INTRO
+51	57	N-WASP	protein	Following their release, the C-terminal regions of N-WASP are free to interact with G-actin and a known nucleator of actin assembly, the Arp2/3 complex.	INTRO
+84	91	G-actin	protein_type	Following their release, the C-terminal regions of N-WASP are free to interact with G-actin and a known nucleator of actin assembly, the Arp2/3 complex.	INTRO
+137	143	Arp2/3	complex_assembly	Following their release, the C-terminal regions of N-WASP are free to interact with G-actin and a known nucleator of actin assembly, the Arp2/3 complex.	INTRO
+18	23	TOCA1	protein	The importance of TOCA1 in actin polymerization has been demonstrated in a range of in vitro and in vivo studies, but the exact role of TOCA1 in the many pathways involving actin assembly remains unclear.	INTRO
+136	141	TOCA1	protein	The importance of TOCA1 in actin polymerization has been demonstrated in a range of in vitro and in vivo studies, but the exact role of TOCA1 in the many pathways involving actin assembly remains unclear.	INTRO
+32	37	TOCA1	protein	The most widely studied role of TOCA1 is in membrane invagination and endocytosis, although it has also been implicated in filopodia formation, neurite elongation, transcriptional reprogramming via nuclear actin, and interaction with ZO-1 at tight junctions.	INTRO
+234	238	ZO-1	protein	The most widely studied role of TOCA1 is in membrane invagination and endocytosis, although it has also been implicated in filopodia formation, neurite elongation, transcriptional reprogramming via nuclear actin, and interaction with ZO-1 at tight junctions.	INTRO
+0	5	TOCA1	protein	TOCA1 comprises an N-terminal F-BAR domain, a central homology region 1 (HR1) domain, and a C-terminal SH3 domain.	INTRO
+30	35	F-BAR	structure_element	TOCA1 comprises an N-terminal F-BAR domain, a central homology region 1 (HR1) domain, and a C-terminal SH3 domain.	INTRO
+46	71	central homology region 1	structure_element	TOCA1 comprises an N-terminal F-BAR domain, a central homology region 1 (HR1) domain, and a C-terminal SH3 domain.	INTRO
+73	76	HR1	structure_element	TOCA1 comprises an N-terminal F-BAR domain, a central homology region 1 (HR1) domain, and a C-terminal SH3 domain.	INTRO
+103	106	SH3	structure_element	TOCA1 comprises an N-terminal F-BAR domain, a central homology region 1 (HR1) domain, and a C-terminal SH3 domain.	INTRO
+4	9	F-BAR	structure_element	The F-BAR domain is a known dimerization, membrane-binding, and membrane-deforming module found in a number of cell signaling proteins.	INTRO
+4	9	TOCA1	protein	The TOCA1 SH3 domain has many known binding partners, including N-WASP and dynamin.	INTRO
+10	13	SH3	structure_element	The TOCA1 SH3 domain has many known binding partners, including N-WASP and dynamin.	INTRO
+64	70	N-WASP	protein	The TOCA1 SH3 domain has many known binding partners, including N-WASP and dynamin.	INTRO
+75	82	dynamin	protein	The TOCA1 SH3 domain has many known binding partners, including N-WASP and dynamin.	INTRO
+4	7	HR1	structure_element	The HR1 domain has been directly implicated in the interaction between TOCA1 and Cdc42, representing the first Cdc42-HR1 domain interaction to be identified.	INTRO
+71	76	TOCA1	protein	The HR1 domain has been directly implicated in the interaction between TOCA1 and Cdc42, representing the first Cdc42-HR1 domain interaction to be identified.	INTRO
+81	86	Cdc42	protein	The HR1 domain has been directly implicated in the interaction between TOCA1 and Cdc42, representing the first Cdc42-HR1 domain interaction to be identified.	INTRO
+111	116	Cdc42	protein	The HR1 domain has been directly implicated in the interaction between TOCA1 and Cdc42, representing the first Cdc42-HR1 domain interaction to be identified.	INTRO
+117	120	HR1	structure_element	The HR1 domain has been directly implicated in the interaction between TOCA1 and Cdc42, representing the first Cdc42-HR1 domain interaction to be identified.	INTRO
+6	9	HR1	structure_element	Other HR1 domains studied so far, including those from the PRK family, have been found to bind their cognate Rho family G protein-binding partner with high specificity and affinities in the nanomolar range.	INTRO
+59	69	PRK family	protein_type	Other HR1 domains studied so far, including those from the PRK family, have been found to bind their cognate Rho family G protein-binding partner with high specificity and affinities in the nanomolar range.	INTRO
+120	129	G protein	protein_type	Other HR1 domains studied so far, including those from the PRK family, have been found to bind their cognate Rho family G protein-binding partner with high specificity and affinities in the nanomolar range.	INTRO
+172	182	affinities	evidence	Other HR1 domains studied so far, including those from the PRK family, have been found to bind their cognate Rho family G protein-binding partner with high specificity and affinities in the nanomolar range.	INTRO
+4	14	structures	evidence	The structures of the PRK1 HR1a domain in complex with RhoA and the HR1b domain in complex with Rac1 show that the HR1 domain comprises an anti-parallel coiled-coil that interacts with its G protein binding partner via both helices.	INTRO
+22	26	PRK1	protein	The structures of the PRK1 HR1a domain in complex with RhoA and the HR1b domain in complex with Rac1 show that the HR1 domain comprises an anti-parallel coiled-coil that interacts with its G protein binding partner via both helices.	INTRO
+27	31	HR1a	structure_element	The structures of the PRK1 HR1a domain in complex with RhoA and the HR1b domain in complex with Rac1 show that the HR1 domain comprises an anti-parallel coiled-coil that interacts with its G protein binding partner via both helices.	INTRO
+42	54	complex with	protein_state	The structures of the PRK1 HR1a domain in complex with RhoA and the HR1b domain in complex with Rac1 show that the HR1 domain comprises an anti-parallel coiled-coil that interacts with its G protein binding partner via both helices.	INTRO
+55	59	RhoA	protein	The structures of the PRK1 HR1a domain in complex with RhoA and the HR1b domain in complex with Rac1 show that the HR1 domain comprises an anti-parallel coiled-coil that interacts with its G protein binding partner via both helices.	INTRO
+68	72	HR1b	structure_element	The structures of the PRK1 HR1a domain in complex with RhoA and the HR1b domain in complex with Rac1 show that the HR1 domain comprises an anti-parallel coiled-coil that interacts with its G protein binding partner via both helices.	INTRO
+83	95	complex with	protein_state	The structures of the PRK1 HR1a domain in complex with RhoA and the HR1b domain in complex with Rac1 show that the HR1 domain comprises an anti-parallel coiled-coil that interacts with its G protein binding partner via both helices.	INTRO
+96	100	Rac1	protein	The structures of the PRK1 HR1a domain in complex with RhoA and the HR1b domain in complex with Rac1 show that the HR1 domain comprises an anti-parallel coiled-coil that interacts with its G protein binding partner via both helices.	INTRO
+115	118	HR1	structure_element	The structures of the PRK1 HR1a domain in complex with RhoA and the HR1b domain in complex with Rac1 show that the HR1 domain comprises an anti-parallel coiled-coil that interacts with its G protein binding partner via both helices.	INTRO
+139	164	anti-parallel coiled-coil	structure_element	The structures of the PRK1 HR1a domain in complex with RhoA and the HR1b domain in complex with Rac1 show that the HR1 domain comprises an anti-parallel coiled-coil that interacts with its G protein binding partner via both helices.	INTRO
+189	198	G protein	protein_type	The structures of the PRK1 HR1a domain in complex with RhoA and the HR1b domain in complex with Rac1 show that the HR1 domain comprises an anti-parallel coiled-coil that interacts with its G protein binding partner via both helices.	INTRO
+224	231	helices	structure_element	The structures of the PRK1 HR1a domain in complex with RhoA and the HR1b domain in complex with Rac1 show that the HR1 domain comprises an anti-parallel coiled-coil that interacts with its G protein binding partner via both helices.	INTRO
+12	36	G protein switch regions	site	Both of the G protein switch regions are involved in the interaction.	INTRO
+4	20	coiled-coil fold	structure_element	The coiled-coil fold is shared by the HR1 domain of the TOCA family protein, CIP4, and, based on sequence homology, by TOCA1 itself.	INTRO
+38	41	HR1	structure_element	The coiled-coil fold is shared by the HR1 domain of the TOCA family protein, CIP4, and, based on sequence homology, by TOCA1 itself.	INTRO
+56	75	TOCA family protein	protein_type	The coiled-coil fold is shared by the HR1 domain of the TOCA family protein, CIP4, and, based on sequence homology, by TOCA1 itself.	INTRO
+77	81	CIP4	protein	The coiled-coil fold is shared by the HR1 domain of the TOCA family protein, CIP4, and, based on sequence homology, by TOCA1 itself.	INTRO
+119	124	TOCA1	protein	The coiled-coil fold is shared by the HR1 domain of the TOCA family protein, CIP4, and, based on sequence homology, by TOCA1 itself.	INTRO
+6	9	HR1	structure_element	These HR1 domains, however, show specificity for Cdc42, rather than RhoA or Rac1.	INTRO
+49	54	Cdc42	protein	These HR1 domains, however, show specificity for Cdc42, rather than RhoA or Rac1.	INTRO
+68	72	RhoA	protein	These HR1 domains, however, show specificity for Cdc42, rather than RhoA or Rac1.	INTRO
+76	80	Rac1	protein	These HR1 domains, however, show specificity for Cdc42, rather than RhoA or Rac1.	INTRO
+14	17	HR1	structure_element	How different HR1 domain proteins distinguish their specific G protein partners remains only partially understood, and structural characterization of a novel G protein-HR1 domain interaction would add to the growing body of information pertaining to these protein complexes.	INTRO
+61	70	G protein	protein_type	How different HR1 domain proteins distinguish their specific G protein partners remains only partially understood, and structural characterization of a novel G protein-HR1 domain interaction would add to the growing body of information pertaining to these protein complexes.	INTRO
+158	167	G protein	protein_type	How different HR1 domain proteins distinguish their specific G protein partners remains only partially understood, and structural characterization of a novel G protein-HR1 domain interaction would add to the growing body of information pertaining to these protein complexes.	INTRO
+168	171	HR1	structure_element	How different HR1 domain proteins distinguish their specific G protein partners remains only partially understood, and structural characterization of a novel G protein-HR1 domain interaction would add to the growing body of information pertaining to these protein complexes.	INTRO
+64	69	TOCA1	protein	Furthermore, the biological function of the interaction between TOCA1 and Cdc42 remains poorly understood, and so far there has been no biophysical or structural insight.	INTRO
+74	79	Cdc42	protein	Furthermore, the biological function of the interaction between TOCA1 and Cdc42 remains poorly understood, and so far there has been no biophysical or structural insight.	INTRO
+20	25	TOCA1	protein	The interactions of TOCA1 and N-WASP with Cdc42 as well as with each other have raised questions as to whether the two Cdc42 effectors can interact with a single molecule of Cdc42 simultaneously.	INTRO
+30	36	N-WASP	protein	The interactions of TOCA1 and N-WASP with Cdc42 as well as with each other have raised questions as to whether the two Cdc42 effectors can interact with a single molecule of Cdc42 simultaneously.	INTRO
+42	47	Cdc42	protein	The interactions of TOCA1 and N-WASP with Cdc42 as well as with each other have raised questions as to whether the two Cdc42 effectors can interact with a single molecule of Cdc42 simultaneously.	INTRO
+119	124	Cdc42	protein	The interactions of TOCA1 and N-WASP with Cdc42 as well as with each other have raised questions as to whether the two Cdc42 effectors can interact with a single molecule of Cdc42 simultaneously.	INTRO
+174	179	Cdc42	protein	The interactions of TOCA1 and N-WASP with Cdc42 as well as with each other have raised questions as to whether the two Cdc42 effectors can interact with a single molecule of Cdc42 simultaneously.	INTRO
+53	58	Cdc42	protein	There is some evidence for a ternary complex between Cdc42, N-WASP, and TOCA1, but there was no direct demonstration of simultaneous contacts between the two effectors and a single molecule of Cdc42.	INTRO
+60	66	N-WASP	protein	There is some evidence for a ternary complex between Cdc42, N-WASP, and TOCA1, but there was no direct demonstration of simultaneous contacts between the two effectors and a single molecule of Cdc42.	INTRO
+72	77	TOCA1	protein	There is some evidence for a ternary complex between Cdc42, N-WASP, and TOCA1, but there was no direct demonstration of simultaneous contacts between the two effectors and a single molecule of Cdc42.	INTRO
+193	198	Cdc42	protein	There is some evidence for a ternary complex between Cdc42, N-WASP, and TOCA1, but there was no direct demonstration of simultaneous contacts between the two effectors and a single molecule of Cdc42.	INTRO
+52	62	structures	evidence	Nonetheless, the substantial difference between the structures of the G protein-binding regions of the two effectors is intriguing and implies that they bind to Cdc42 quite differently, providing motivation for investigating the possibility that Cdc42 can bind both effectors concurrently.	INTRO
+70	95	G protein-binding regions	site	Nonetheless, the substantial difference between the structures of the G protein-binding regions of the two effectors is intriguing and implies that they bind to Cdc42 quite differently, providing motivation for investigating the possibility that Cdc42 can bind both effectors concurrently.	INTRO
+161	166	Cdc42	protein	Nonetheless, the substantial difference between the structures of the G protein-binding regions of the two effectors is intriguing and implies that they bind to Cdc42 quite differently, providing motivation for investigating the possibility that Cdc42 can bind both effectors concurrently.	INTRO
+246	251	Cdc42	protein	Nonetheless, the substantial difference between the structures of the G protein-binding regions of the two effectors is intriguing and implies that they bind to Cdc42 quite differently, providing motivation for investigating the possibility that Cdc42 can bind both effectors concurrently.	INTRO
+0	4	WASP	protein_type	WASP interacts with Cdc42 via a conserved, unstructured binding motif known as the Cdc42- and Rac-interactive binding region (CRIB), which forms an intermolecular β-sheet, expanding the anti-parallel β2 and β3 strands of Cdc42.	INTRO
+20	25	Cdc42	protein	WASP interacts with Cdc42 via a conserved, unstructured binding motif known as the Cdc42- and Rac-interactive binding region (CRIB), which forms an intermolecular β-sheet, expanding the anti-parallel β2 and β3 strands of Cdc42.	INTRO
+32	41	conserved	protein_state	WASP interacts with Cdc42 via a conserved, unstructured binding motif known as the Cdc42- and Rac-interactive binding region (CRIB), which forms an intermolecular β-sheet, expanding the anti-parallel β2 and β3 strands of Cdc42.	INTRO
+43	69	unstructured binding motif	structure_element	WASP interacts with Cdc42 via a conserved, unstructured binding motif known as the Cdc42- and Rac-interactive binding region (CRIB), which forms an intermolecular β-sheet, expanding the anti-parallel β2 and β3 strands of Cdc42.	INTRO
+83	124	Cdc42- and Rac-interactive binding region	structure_element	WASP interacts with Cdc42 via a conserved, unstructured binding motif known as the Cdc42- and Rac-interactive binding region (CRIB), which forms an intermolecular β-sheet, expanding the anti-parallel β2 and β3 strands of Cdc42.	INTRO
+126	130	CRIB	structure_element	WASP interacts with Cdc42 via a conserved, unstructured binding motif known as the Cdc42- and Rac-interactive binding region (CRIB), which forms an intermolecular β-sheet, expanding the anti-parallel β2 and β3 strands of Cdc42.	INTRO
+148	170	intermolecular β-sheet	structure_element	WASP interacts with Cdc42 via a conserved, unstructured binding motif known as the Cdc42- and Rac-interactive binding region (CRIB), which forms an intermolecular β-sheet, expanding the anti-parallel β2 and β3 strands of Cdc42.	INTRO
+200	217	β2 and β3 strands	structure_element	WASP interacts with Cdc42 via a conserved, unstructured binding motif known as the Cdc42- and Rac-interactive binding region (CRIB), which forms an intermolecular β-sheet, expanding the anti-parallel β2 and β3 strands of Cdc42.	INTRO
+221	226	Cdc42	protein	WASP interacts with Cdc42 via a conserved, unstructured binding motif known as the Cdc42- and Rac-interactive binding region (CRIB), which forms an intermolecular β-sheet, expanding the anti-parallel β2 and β3 strands of Cdc42.	INTRO
+17	37	TOCA family proteins	protein_type	In contrast, the TOCA family proteins are thought to interact via the HR1 domain, which may form a triple coiled-coil with switch II of Rac1, like the HR1b domain of PRK1.	INTRO
+70	73	HR1	structure_element	In contrast, the TOCA family proteins are thought to interact via the HR1 domain, which may form a triple coiled-coil with switch II of Rac1, like the HR1b domain of PRK1.	INTRO
+99	117	triple coiled-coil	structure_element	In contrast, the TOCA family proteins are thought to interact via the HR1 domain, which may form a triple coiled-coil with switch II of Rac1, like the HR1b domain of PRK1.	INTRO
+123	132	switch II	site	In contrast, the TOCA family proteins are thought to interact via the HR1 domain, which may form a triple coiled-coil with switch II of Rac1, like the HR1b domain of PRK1.	INTRO
+136	140	Rac1	protein	In contrast, the TOCA family proteins are thought to interact via the HR1 domain, which may form a triple coiled-coil with switch II of Rac1, like the HR1b domain of PRK1.	INTRO
+151	155	HR1b	structure_element	In contrast, the TOCA family proteins are thought to interact via the HR1 domain, which may form a triple coiled-coil with switch II of Rac1, like the HR1b domain of PRK1.	INTRO
+166	170	PRK1	protein	In contrast, the TOCA family proteins are thought to interact via the HR1 domain, which may form a triple coiled-coil with switch II of Rac1, like the HR1b domain of PRK1.	INTRO
+21	33	solution NMR	experimental_method	Here, we present the solution NMR structure of the HR1 domain of TOCA1, providing the first structural data for this protein.	INTRO
+34	43	structure	evidence	Here, we present the solution NMR structure of the HR1 domain of TOCA1, providing the first structural data for this protein.	INTRO
+51	54	HR1	structure_element	Here, we present the solution NMR structure of the HR1 domain of TOCA1, providing the first structural data for this protein.	INTRO
+65	70	TOCA1	protein	Here, we present the solution NMR structure of the HR1 domain of TOCA1, providing the first structural data for this protein.	INTRO
+92	107	structural data	evidence	Here, we present the solution NMR structure of the HR1 domain of TOCA1, providing the first structural data for this protein.	INTRO
+50	54	TOCA	protein_type	We also present data pertaining to binding of the TOCA HR1 domain to Cdc42, which is the first biophysical description of an HR1 domain binding this particular Rho family small G protein.	INTRO
+55	58	HR1	structure_element	We also present data pertaining to binding of the TOCA HR1 domain to Cdc42, which is the first biophysical description of an HR1 domain binding this particular Rho family small G protein.	INTRO
+69	74	Cdc42	protein	We also present data pertaining to binding of the TOCA HR1 domain to Cdc42, which is the first biophysical description of an HR1 domain binding this particular Rho family small G protein.	INTRO
+125	128	HR1	structure_element	We also present data pertaining to binding of the TOCA HR1 domain to Cdc42, which is the first biophysical description of an HR1 domain binding this particular Rho family small G protein.	INTRO
+160	186	Rho family small G protein	protein_type	We also present data pertaining to binding of the TOCA HR1 domain to Cdc42, which is the first biophysical description of an HR1 domain binding this particular Rho family small G protein.	INTRO
+62	67	Cdc42	protein	Finally, we investigate the potential ternary complex between Cdc42 and the G protein-binding regions of TOCA1 and N-WASP, contributing to our understanding of G protein-effector interactions as well as the roles of Cdc42, N-WASP, and TOCA1 in the pathways that govern actin dynamics.	INTRO
+76	101	G protein-binding regions	site	Finally, we investigate the potential ternary complex between Cdc42 and the G protein-binding regions of TOCA1 and N-WASP, contributing to our understanding of G protein-effector interactions as well as the roles of Cdc42, N-WASP, and TOCA1 in the pathways that govern actin dynamics.	INTRO
+105	110	TOCA1	protein	Finally, we investigate the potential ternary complex between Cdc42 and the G protein-binding regions of TOCA1 and N-WASP, contributing to our understanding of G protein-effector interactions as well as the roles of Cdc42, N-WASP, and TOCA1 in the pathways that govern actin dynamics.	INTRO
+115	121	N-WASP	protein	Finally, we investigate the potential ternary complex between Cdc42 and the G protein-binding regions of TOCA1 and N-WASP, contributing to our understanding of G protein-effector interactions as well as the roles of Cdc42, N-WASP, and TOCA1 in the pathways that govern actin dynamics.	INTRO
+160	169	G protein	protein_type	Finally, we investigate the potential ternary complex between Cdc42 and the G protein-binding regions of TOCA1 and N-WASP, contributing to our understanding of G protein-effector interactions as well as the roles of Cdc42, N-WASP, and TOCA1 in the pathways that govern actin dynamics.	INTRO
+216	221	Cdc42	protein	Finally, we investigate the potential ternary complex between Cdc42 and the G protein-binding regions of TOCA1 and N-WASP, contributing to our understanding of G protein-effector interactions as well as the roles of Cdc42, N-WASP, and TOCA1 in the pathways that govern actin dynamics.	INTRO
+223	229	N-WASP	protein	Finally, we investigate the potential ternary complex between Cdc42 and the G protein-binding regions of TOCA1 and N-WASP, contributing to our understanding of G protein-effector interactions as well as the roles of Cdc42, N-WASP, and TOCA1 in the pathways that govern actin dynamics.	INTRO
+235	240	TOCA1	protein	Finally, we investigate the potential ternary complex between Cdc42 and the G protein-binding regions of TOCA1 and N-WASP, contributing to our understanding of G protein-effector interactions as well as the roles of Cdc42, N-WASP, and TOCA1 in the pathways that govern actin dynamics.	INTRO
+0	5	Cdc42	protein	Cdc42-TOCA1 Binding	RESULTS
+6	11	TOCA1	protein	Cdc42-TOCA1 Binding	RESULTS
+0	5	TOCA1	protein	TOCA1 was identified in Xenopus extracts as a protein necessary for Cdc42-dependent actin assembly and was shown to bind to Cdc42·GTPγS but not to Cdc42·GDP or to Rac1 and RhoA. Given its homology to other Rho family binding modules, it is likely that the HR1 domain of TOCA1 is sufficient to bind Cdc42.	RESULTS
+24	31	Xenopus	taxonomy_domain	TOCA1 was identified in Xenopus extracts as a protein necessary for Cdc42-dependent actin assembly and was shown to bind to Cdc42·GTPγS but not to Cdc42·GDP or to Rac1 and RhoA. Given its homology to other Rho family binding modules, it is likely that the HR1 domain of TOCA1 is sufficient to bind Cdc42.	RESULTS
+68	73	Cdc42	protein	TOCA1 was identified in Xenopus extracts as a protein necessary for Cdc42-dependent actin assembly and was shown to bind to Cdc42·GTPγS but not to Cdc42·GDP or to Rac1 and RhoA. Given its homology to other Rho family binding modules, it is likely that the HR1 domain of TOCA1 is sufficient to bind Cdc42.	RESULTS
+124	135	Cdc42·GTPγS	complex_assembly	TOCA1 was identified in Xenopus extracts as a protein necessary for Cdc42-dependent actin assembly and was shown to bind to Cdc42·GTPγS but not to Cdc42·GDP or to Rac1 and RhoA. Given its homology to other Rho family binding modules, it is likely that the HR1 domain of TOCA1 is sufficient to bind Cdc42.	RESULTS
+147	156	Cdc42·GDP	complex_assembly	TOCA1 was identified in Xenopus extracts as a protein necessary for Cdc42-dependent actin assembly and was shown to bind to Cdc42·GTPγS but not to Cdc42·GDP or to Rac1 and RhoA. Given its homology to other Rho family binding modules, it is likely that the HR1 domain of TOCA1 is sufficient to bind Cdc42.	RESULTS
+163	167	Rac1	protein	TOCA1 was identified in Xenopus extracts as a protein necessary for Cdc42-dependent actin assembly and was shown to bind to Cdc42·GTPγS but not to Cdc42·GDP or to Rac1 and RhoA. Given its homology to other Rho family binding modules, it is likely that the HR1 domain of TOCA1 is sufficient to bind Cdc42.	RESULTS
+172	176	RhoA	protein	TOCA1 was identified in Xenopus extracts as a protein necessary for Cdc42-dependent actin assembly and was shown to bind to Cdc42·GTPγS but not to Cdc42·GDP or to Rac1 and RhoA. Given its homology to other Rho family binding modules, it is likely that the HR1 domain of TOCA1 is sufficient to bind Cdc42.	RESULTS
+206	232	Rho family binding modules	site	TOCA1 was identified in Xenopus extracts as a protein necessary for Cdc42-dependent actin assembly and was shown to bind to Cdc42·GTPγS but not to Cdc42·GDP or to Rac1 and RhoA. Given its homology to other Rho family binding modules, it is likely that the HR1 domain of TOCA1 is sufficient to bind Cdc42.	RESULTS
+256	259	HR1	structure_element	TOCA1 was identified in Xenopus extracts as a protein necessary for Cdc42-dependent actin assembly and was shown to bind to Cdc42·GTPγS but not to Cdc42·GDP or to Rac1 and RhoA. Given its homology to other Rho family binding modules, it is likely that the HR1 domain of TOCA1 is sufficient to bind Cdc42.	RESULTS
+270	275	TOCA1	protein	TOCA1 was identified in Xenopus extracts as a protein necessary for Cdc42-dependent actin assembly and was shown to bind to Cdc42·GTPγS but not to Cdc42·GDP or to Rac1 and RhoA. Given its homology to other Rho family binding modules, it is likely that the HR1 domain of TOCA1 is sufficient to bind Cdc42.	RESULTS
+298	303	Cdc42	protein	TOCA1 was identified in Xenopus extracts as a protein necessary for Cdc42-dependent actin assembly and was shown to bind to Cdc42·GTPγS but not to Cdc42·GDP or to Rac1 and RhoA. Given its homology to other Rho family binding modules, it is likely that the HR1 domain of TOCA1 is sufficient to bind Cdc42.	RESULTS
+4	14	C. elegans	species	The C. elegans TOCA1 orthologues also bind to Cdc42 via their consensus HR1 domain.	RESULTS
+15	20	TOCA1	protein	The C. elegans TOCA1 orthologues also bind to Cdc42 via their consensus HR1 domain.	RESULTS
+46	51	Cdc42	protein	The C. elegans TOCA1 orthologues also bind to Cdc42 via their consensus HR1 domain.	RESULTS
+72	75	HR1	structure_element	The C. elegans TOCA1 orthologues also bind to Cdc42 via their consensus HR1 domain.	RESULTS
+4	7	HR1	structure_element	The HR1 domains from the PRK family bind their G protein partners with a high affinity, exhibiting a range of submicromolar dissociation constants (Kd) as low as 26 nm.	RESULTS
+25	35	PRK family	protein_type	The HR1 domains from the PRK family bind their G protein partners with a high affinity, exhibiting a range of submicromolar dissociation constants (Kd) as low as 26 nm.	RESULTS
+47	56	G protein	protein_type	The HR1 domains from the PRK family bind their G protein partners with a high affinity, exhibiting a range of submicromolar dissociation constants (Kd) as low as 26 nm.	RESULTS
+124	146	dissociation constants	evidence	The HR1 domains from the PRK family bind their G protein partners with a high affinity, exhibiting a range of submicromolar dissociation constants (Kd) as low as 26 nm.	RESULTS
+148	150	Kd	evidence	The HR1 domains from the PRK family bind their G protein partners with a high affinity, exhibiting a range of submicromolar dissociation constants (Kd) as low as 26 nm.	RESULTS
+2	4	Kd	evidence	A Kd in the nanomolar range was therefore expected for the interaction of the TOCA1 HR1 domain with Cdc42.	RESULTS
+78	83	TOCA1	protein	A Kd in the nanomolar range was therefore expected for the interaction of the TOCA1 HR1 domain with Cdc42.	RESULTS
+84	87	HR1	structure_element	A Kd in the nanomolar range was therefore expected for the interaction of the TOCA1 HR1 domain with Cdc42.	RESULTS
+100	105	Cdc42	protein	A Kd in the nanomolar range was therefore expected for the interaction of the TOCA1 HR1 domain with Cdc42.	RESULTS
+16	29	X. tropicalis	species	We generated an X. tropicalis TOCA1 HR1 domain construct encompassing residues 330–426.	RESULTS
+30	35	TOCA1	protein	We generated an X. tropicalis TOCA1 HR1 domain construct encompassing residues 330–426.	RESULTS
+36	39	HR1	structure_element	We generated an X. tropicalis TOCA1 HR1 domain construct encompassing residues 330–426.	RESULTS
+79	86	330–426	residue_range	We generated an X. tropicalis TOCA1 HR1 domain construct encompassing residues 330–426.	RESULTS
+35	38	HR1	structure_element	This region comprises the complete HR1 domain based on secondary structure predictions and sequence alignments with another TOCA family member, CIP4, whose structure has been determined.	RESULTS
+91	110	sequence alignments	experimental_method	This region comprises the complete HR1 domain based on secondary structure predictions and sequence alignments with another TOCA family member, CIP4, whose structure has been determined.	RESULTS
+124	135	TOCA family	protein_type	This region comprises the complete HR1 domain based on secondary structure predictions and sequence alignments with another TOCA family member, CIP4, whose structure has been determined.	RESULTS
+144	148	CIP4	protein	This region comprises the complete HR1 domain based on secondary structure predictions and sequence alignments with another TOCA family member, CIP4, whose structure has been determined.	RESULTS
+156	165	structure	evidence	This region comprises the complete HR1 domain based on secondary structure predictions and sequence alignments with another TOCA family member, CIP4, whose structure has been determined.	RESULTS
+24	37	[3H]GTP·Cdc42	complex_assembly	The interaction between [3H]GTP·Cdc42 and a C-terminally His-tagged TOCA1 HR1 domain construct was investigated using SPA.	RESULTS
+57	67	His-tagged	protein_state	The interaction between [3H]GTP·Cdc42 and a C-terminally His-tagged TOCA1 HR1 domain construct was investigated using SPA.	RESULTS
+68	73	TOCA1	protein	The interaction between [3H]GTP·Cdc42 and a C-terminally His-tagged TOCA1 HR1 domain construct was investigated using SPA.	RESULTS
+74	77	HR1	structure_element	The interaction between [3H]GTP·Cdc42 and a C-terminally His-tagged TOCA1 HR1 domain construct was investigated using SPA.	RESULTS
+118	121	SPA	experimental_method	The interaction between [3H]GTP·Cdc42 and a C-terminally His-tagged TOCA1 HR1 domain construct was investigated using SPA.	RESULTS
+4	20	binding isotherm	evidence	The binding isotherm for the interaction is shown in Fig. 1A, together with the Cdc42-PAK interaction as a positive control.	RESULTS
+80	85	Cdc42	protein	The binding isotherm for the interaction is shown in Fig. 1A, together with the Cdc42-PAK interaction as a positive control.	RESULTS
+86	89	PAK	protein	The binding isotherm for the interaction is shown in Fig. 1A, together with the Cdc42-PAK interaction as a positive control.	RESULTS
+15	20	TOCA1	protein	The binding of TOCA1 HR1 to Cdc42 was unexpectedly weak, with a Kd of >1 μm.	RESULTS
+21	24	HR1	structure_element	The binding of TOCA1 HR1 to Cdc42 was unexpectedly weak, with a Kd of >1 μm.	RESULTS
+28	33	Cdc42	protein	The binding of TOCA1 HR1 to Cdc42 was unexpectedly weak, with a Kd of >1 μm.	RESULTS
+64	66	Kd	evidence	The binding of TOCA1 HR1 to Cdc42 was unexpectedly weak, with a Kd of >1 μm.	RESULTS
+36	38	Kd	evidence	It was not possible to estimate the Kd more accurately using direct SPA experiments, because saturation could not be reached due to nonspecific signal at higher protein concentrations.	RESULTS
+68	71	SPA	experimental_method	It was not possible to estimate the Kd more accurately using direct SPA experiments, because saturation could not be reached due to nonspecific signal at higher protein concentrations.	RESULTS
+4	9	TOCA1	protein	The TOCA1 HR1-Cdc42 interaction is low affinity.	FIG
+10	13	HR1	structure_element	The TOCA1 HR1-Cdc42 interaction is low affinity.	FIG
+14	19	Cdc42	protein	The TOCA1 HR1-Cdc42 interaction is low affinity.	FIG
+24	45	direct binding assays	experimental_method	 A, curves derived from direct binding assays in which the indicated concentrations of Cdc42Δ7Q61L·[3H]GTP were incubated with 30 nm GST-PAK or HR1-His6 in SPAs.	FIG
+87	106	Cdc42Δ7Q61L·[3H]GTP	complex_assembly	 A, curves derived from direct binding assays in which the indicated concentrations of Cdc42Δ7Q61L·[3H]GTP were incubated with 30 nm GST-PAK or HR1-His6 in SPAs.	FIG
+112	121	incubated	experimental_method	 A, curves derived from direct binding assays in which the indicated concentrations of Cdc42Δ7Q61L·[3H]GTP were incubated with 30 nm GST-PAK or HR1-His6 in SPAs.	FIG
+133	140	GST-PAK	mutant	 A, curves derived from direct binding assays in which the indicated concentrations of Cdc42Δ7Q61L·[3H]GTP were incubated with 30 nm GST-PAK or HR1-His6 in SPAs.	FIG
+144	152	HR1-His6	mutant	 A, curves derived from direct binding assays in which the indicated concentrations of Cdc42Δ7Q61L·[3H]GTP were incubated with 30 nm GST-PAK or HR1-His6 in SPAs.	FIG
+156	160	SPAs	experimental_method	 A, curves derived from direct binding assays in which the indicated concentrations of Cdc42Δ7Q61L·[3H]GTP were incubated with 30 nm GST-PAK or HR1-His6 in SPAs.	FIG
+4	7	SPA	experimental_method	The SPA signal was corrected by subtraction of control data with no GST-PAK or HR1-His6.	FIG
+68	75	GST-PAK	mutant	The SPA signal was corrected by subtraction of control data with no GST-PAK or HR1-His6.	FIG
+79	87	HR1-His6	mutant	The SPA signal was corrected by subtraction of control data with no GST-PAK or HR1-His6.	FIG
+26	42	binding isotherm	evidence	The data were fitted to a binding isotherm to give an apparent Kd and are expressed as a percentage of the maximum signal; B and C, competition SPA experiments were carried out with the indicated concentrations of ACK GBD (B) or HR1 domain (C) titrated into 30 nm GST-ACK and either 30 nm Cdc42Δ7Q61L·[3H]GTP or full-length Cdc42Q61L·[3H]GTP.	FIG
+63	65	Kd	evidence	The data were fitted to a binding isotherm to give an apparent Kd and are expressed as a percentage of the maximum signal; B and C, competition SPA experiments were carried out with the indicated concentrations of ACK GBD (B) or HR1 domain (C) titrated into 30 nm GST-ACK and either 30 nm Cdc42Δ7Q61L·[3H]GTP or full-length Cdc42Q61L·[3H]GTP.	FIG
+132	147	competition SPA	experimental_method	The data were fitted to a binding isotherm to give an apparent Kd and are expressed as a percentage of the maximum signal; B and C, competition SPA experiments were carried out with the indicated concentrations of ACK GBD (B) or HR1 domain (C) titrated into 30 nm GST-ACK and either 30 nm Cdc42Δ7Q61L·[3H]GTP or full-length Cdc42Q61L·[3H]GTP.	FIG
+214	217	ACK	protein	The data were fitted to a binding isotherm to give an apparent Kd and are expressed as a percentage of the maximum signal; B and C, competition SPA experiments were carried out with the indicated concentrations of ACK GBD (B) or HR1 domain (C) titrated into 30 nm GST-ACK and either 30 nm Cdc42Δ7Q61L·[3H]GTP or full-length Cdc42Q61L·[3H]GTP.	FIG
+218	221	GBD	structure_element	The data were fitted to a binding isotherm to give an apparent Kd and are expressed as a percentage of the maximum signal; B and C, competition SPA experiments were carried out with the indicated concentrations of ACK GBD (B) or HR1 domain (C) titrated into 30 nm GST-ACK and either 30 nm Cdc42Δ7Q61L·[3H]GTP or full-length Cdc42Q61L·[3H]GTP.	FIG
+229	232	HR1	structure_element	The data were fitted to a binding isotherm to give an apparent Kd and are expressed as a percentage of the maximum signal; B and C, competition SPA experiments were carried out with the indicated concentrations of ACK GBD (B) or HR1 domain (C) titrated into 30 nm GST-ACK and either 30 nm Cdc42Δ7Q61L·[3H]GTP or full-length Cdc42Q61L·[3H]GTP.	FIG
+244	252	titrated	experimental_method	The data were fitted to a binding isotherm to give an apparent Kd and are expressed as a percentage of the maximum signal; B and C, competition SPA experiments were carried out with the indicated concentrations of ACK GBD (B) or HR1 domain (C) titrated into 30 nm GST-ACK and either 30 nm Cdc42Δ7Q61L·[3H]GTP or full-length Cdc42Q61L·[3H]GTP.	FIG
+264	271	GST-ACK	mutant	The data were fitted to a binding isotherm to give an apparent Kd and are expressed as a percentage of the maximum signal; B and C, competition SPA experiments were carried out with the indicated concentrations of ACK GBD (B) or HR1 domain (C) titrated into 30 nm GST-ACK and either 30 nm Cdc42Δ7Q61L·[3H]GTP or full-length Cdc42Q61L·[3H]GTP.	FIG
+289	308	Cdc42Δ7Q61L·[3H]GTP	complex_assembly	The data were fitted to a binding isotherm to give an apparent Kd and are expressed as a percentage of the maximum signal; B and C, competition SPA experiments were carried out with the indicated concentrations of ACK GBD (B) or HR1 domain (C) titrated into 30 nm GST-ACK and either 30 nm Cdc42Δ7Q61L·[3H]GTP or full-length Cdc42Q61L·[3H]GTP.	FIG
+312	323	full-length	protein_state	The data were fitted to a binding isotherm to give an apparent Kd and are expressed as a percentage of the maximum signal; B and C, competition SPA experiments were carried out with the indicated concentrations of ACK GBD (B) or HR1 domain (C) titrated into 30 nm GST-ACK and either 30 nm Cdc42Δ7Q61L·[3H]GTP or full-length Cdc42Q61L·[3H]GTP.	FIG
+324	341	Cdc42Q61L·[3H]GTP	complex_assembly	The data were fitted to a binding isotherm to give an apparent Kd and are expressed as a percentage of the maximum signal; B and C, competition SPA experiments were carried out with the indicated concentrations of ACK GBD (B) or HR1 domain (C) titrated into 30 nm GST-ACK and either 30 nm Cdc42Δ7Q61L·[3H]GTP or full-length Cdc42Q61L·[3H]GTP.	FIG
+4	6	Kd	evidence	The Kd values derived for the ACK GBD with Cdc42Δ7 and full-length Cdc42 were 0.032 ± 0.01 and 0.011 ± 0.01 μm, respectively.	FIG
+30	33	ACK	protein	The Kd values derived for the ACK GBD with Cdc42Δ7 and full-length Cdc42 were 0.032 ± 0.01 and 0.011 ± 0.01 μm, respectively.	FIG
+34	37	GBD	structure_element	The Kd values derived for the ACK GBD with Cdc42Δ7 and full-length Cdc42 were 0.032 ± 0.01 and 0.011 ± 0.01 μm, respectively.	FIG
+43	50	Cdc42Δ7	mutant	The Kd values derived for the ACK GBD with Cdc42Δ7 and full-length Cdc42 were 0.032 ± 0.01 and 0.011 ± 0.01 μm, respectively.	FIG
+55	66	full-length	protein_state	The Kd values derived for the ACK GBD with Cdc42Δ7 and full-length Cdc42 were 0.032 ± 0.01 and 0.011 ± 0.01 μm, respectively.	FIG
+67	72	Cdc42	protein	The Kd values derived for the ACK GBD with Cdc42Δ7 and full-length Cdc42 were 0.032 ± 0.01 and 0.011 ± 0.01 μm, respectively.	FIG
+4	6	Kd	evidence	The Kd values derived for the TOCA1 HR1 with Cdc42Δ7 and full-length Cdc42 were 6.05 ± 1.96 and 5.39 ± 1.69 μm, respectively.	FIG
+30	35	TOCA1	protein	The Kd values derived for the TOCA1 HR1 with Cdc42Δ7 and full-length Cdc42 were 6.05 ± 1.96 and 5.39 ± 1.69 μm, respectively.	FIG
+36	39	HR1	structure_element	The Kd values derived for the TOCA1 HR1 with Cdc42Δ7 and full-length Cdc42 were 6.05 ± 1.96 and 5.39 ± 1.69 μm, respectively.	FIG
+45	52	Cdc42Δ7	mutant	The Kd values derived for the TOCA1 HR1 with Cdc42Δ7 and full-length Cdc42 were 6.05 ± 1.96 and 5.39 ± 1.69 μm, respectively.	FIG
+57	68	full-length	protein_state	The Kd values derived for the TOCA1 HR1 with Cdc42Δ7 and full-length Cdc42 were 6.05 ± 1.96 and 5.39 ± 1.69 μm, respectively.	FIG
+69	74	Cdc42	protein	The Kd values derived for the TOCA1 HR1 with Cdc42Δ7 and full-length Cdc42 were 6.05 ± 1.96 and 5.39 ± 1.69 μm, respectively.	FIG
+100	103	HR1	structure_element	It was possible that the low affinity observed was due to negative effects of immobilization of the HR1 domain, so other methods were employed, which utilized untagged proteins.	RESULTS
+159	167	untagged	protein_state	It was possible that the low affinity observed was due to negative effects of immobilization of the HR1 domain, so other methods were employed, which utilized untagged proteins.	RESULTS
+0	32	Isothermal titration calorimetry	experimental_method	Isothermal titration calorimetry was carried out, but no heat changes were observed at a range of concentrations and temperatures (data not shown), suggesting that the interaction is predominantly entropically driven.	RESULTS
+6	15	G protein	protein_type	Other G protein-HR1 domain interactions have also failed to show heat changes in our hands.7 Infrared interferometry with immobilized Cdc42 was also attempted but was unsuccessful for both TOCA1 HR1 and for the positive control, ACK.	RESULTS
+16	19	HR1	structure_element	Other G protein-HR1 domain interactions have also failed to show heat changes in our hands.7 Infrared interferometry with immobilized Cdc42 was also attempted but was unsuccessful for both TOCA1 HR1 and for the positive control, ACK.	RESULTS
+93	116	Infrared interferometry	experimental_method	Other G protein-HR1 domain interactions have also failed to show heat changes in our hands.7 Infrared interferometry with immobilized Cdc42 was also attempted but was unsuccessful for both TOCA1 HR1 and for the positive control, ACK.	RESULTS
+122	133	immobilized	protein_state	Other G protein-HR1 domain interactions have also failed to show heat changes in our hands.7 Infrared interferometry with immobilized Cdc42 was also attempted but was unsuccessful for both TOCA1 HR1 and for the positive control, ACK.	RESULTS
+134	139	Cdc42	protein	Other G protein-HR1 domain interactions have also failed to show heat changes in our hands.7 Infrared interferometry with immobilized Cdc42 was also attempted but was unsuccessful for both TOCA1 HR1 and for the positive control, ACK.	RESULTS
+189	194	TOCA1	protein	Other G protein-HR1 domain interactions have also failed to show heat changes in our hands.7 Infrared interferometry with immobilized Cdc42 was also attempted but was unsuccessful for both TOCA1 HR1 and for the positive control, ACK.	RESULTS
+195	198	HR1	structure_element	Other G protein-HR1 domain interactions have also failed to show heat changes in our hands.7 Infrared interferometry with immobilized Cdc42 was also attempted but was unsuccessful for both TOCA1 HR1 and for the positive control, ACK.	RESULTS
+229	232	ACK	protein	Other G protein-HR1 domain interactions have also failed to show heat changes in our hands.7 Infrared interferometry with immobilized Cdc42 was also attempted but was unsuccessful for both TOCA1 HR1 and for the positive control, ACK.	RESULTS
+4	12	affinity	evidence	The affinity was therefore determined using competition SPAs.	RESULTS
+44	60	competition SPAs	experimental_method	The affinity was therefore determined using competition SPAs.	RESULTS
+15	25	GST fusion	experimental_method	A complex of a GST fusion of the GBD of ACK, which binds with a high affinity to Cdc42, with radiolabeled [3H]GTP·Cdc42 was preformed, and the effect of increasing concentrations of untagged TOCA1 HR1 domain was examined.	RESULTS
+33	36	GBD	structure_element	A complex of a GST fusion of the GBD of ACK, which binds with a high affinity to Cdc42, with radiolabeled [3H]GTP·Cdc42 was preformed, and the effect of increasing concentrations of untagged TOCA1 HR1 domain was examined.	RESULTS
+40	43	ACK	protein	A complex of a GST fusion of the GBD of ACK, which binds with a high affinity to Cdc42, with radiolabeled [3H]GTP·Cdc42 was preformed, and the effect of increasing concentrations of untagged TOCA1 HR1 domain was examined.	RESULTS
+81	86	Cdc42	protein	A complex of a GST fusion of the GBD of ACK, which binds with a high affinity to Cdc42, with radiolabeled [3H]GTP·Cdc42 was preformed, and the effect of increasing concentrations of untagged TOCA1 HR1 domain was examined.	RESULTS
+106	119	[3H]GTP·Cdc42	complex_assembly	A complex of a GST fusion of the GBD of ACK, which binds with a high affinity to Cdc42, with radiolabeled [3H]GTP·Cdc42 was preformed, and the effect of increasing concentrations of untagged TOCA1 HR1 domain was examined.	RESULTS
+153	178	increasing concentrations	experimental_method	A complex of a GST fusion of the GBD of ACK, which binds with a high affinity to Cdc42, with radiolabeled [3H]GTP·Cdc42 was preformed, and the effect of increasing concentrations of untagged TOCA1 HR1 domain was examined.	RESULTS
+182	190	untagged	protein_state	A complex of a GST fusion of the GBD of ACK, which binds with a high affinity to Cdc42, with radiolabeled [3H]GTP·Cdc42 was preformed, and the effect of increasing concentrations of untagged TOCA1 HR1 domain was examined.	RESULTS
+191	196	TOCA1	protein	A complex of a GST fusion of the GBD of ACK, which binds with a high affinity to Cdc42, with radiolabeled [3H]GTP·Cdc42 was preformed, and the effect of increasing concentrations of untagged TOCA1 HR1 domain was examined.	RESULTS
+197	200	HR1	structure_element	A complex of a GST fusion of the GBD of ACK, which binds with a high affinity to Cdc42, with radiolabeled [3H]GTP·Cdc42 was preformed, and the effect of increasing concentrations of untagged TOCA1 HR1 domain was examined.	RESULTS
+15	22	GST-ACK	mutant	Competition of GST-ACK GBD bound to [3H]GTP·Cdc42 by free ACK GBD was used as a control and to establish the value of background counts when Cdc42 is fully displaced.	RESULTS
+23	26	GBD	structure_element	Competition of GST-ACK GBD bound to [3H]GTP·Cdc42 by free ACK GBD was used as a control and to establish the value of background counts when Cdc42 is fully displaced.	RESULTS
+27	35	bound to	protein_state	Competition of GST-ACK GBD bound to [3H]GTP·Cdc42 by free ACK GBD was used as a control and to establish the value of background counts when Cdc42 is fully displaced.	RESULTS
+36	49	[3H]GTP·Cdc42	complex_assembly	Competition of GST-ACK GBD bound to [3H]GTP·Cdc42 by free ACK GBD was used as a control and to establish the value of background counts when Cdc42 is fully displaced.	RESULTS
+53	57	free	protein_state	Competition of GST-ACK GBD bound to [3H]GTP·Cdc42 by free ACK GBD was used as a control and to establish the value of background counts when Cdc42 is fully displaced.	RESULTS
+58	61	ACK	protein	Competition of GST-ACK GBD bound to [3H]GTP·Cdc42 by free ACK GBD was used as a control and to establish the value of background counts when Cdc42 is fully displaced.	RESULTS
+62	65	GBD	structure_element	Competition of GST-ACK GBD bound to [3H]GTP·Cdc42 by free ACK GBD was used as a control and to establish the value of background counts when Cdc42 is fully displaced.	RESULTS
+141	146	Cdc42	protein	Competition of GST-ACK GBD bound to [3H]GTP·Cdc42 by free ACK GBD was used as a control and to establish the value of background counts when Cdc42 is fully displaced.	RESULTS
+26	42	binding isotherm	evidence	The data were fitted to a binding isotherm describing competition.	RESULTS
+0	4	Free	protein_state	Free ACK competed with itself with an affinity of 32 nm, similar to the value obtained by direct binding of 23 nm.	RESULTS
+5	8	ACK	protein	Free ACK competed with itself with an affinity of 32 nm, similar to the value obtained by direct binding of 23 nm.	RESULTS
+38	46	affinity	evidence	Free ACK competed with itself with an affinity of 32 nm, similar to the value obtained by direct binding of 23 nm.	RESULTS
+90	104	direct binding	experimental_method	Free ACK competed with itself with an affinity of 32 nm, similar to the value obtained by direct binding of 23 nm.	RESULTS
+4	9	TOCA1	protein	The TOCA1 HR1 domain also fully competed with the GST-ACK but bound with an affinity of 6 μm (Fig. 1, B and C), in agreement with the low affinity observed in the direct binding experiments.	RESULTS
+10	13	HR1	structure_element	The TOCA1 HR1 domain also fully competed with the GST-ACK but bound with an affinity of 6 μm (Fig. 1, B and C), in agreement with the low affinity observed in the direct binding experiments.	RESULTS
+50	57	GST-ACK	mutant	The TOCA1 HR1 domain also fully competed with the GST-ACK but bound with an affinity of 6 μm (Fig. 1, B and C), in agreement with the low affinity observed in the direct binding experiments.	RESULTS
+62	67	bound	protein_state	The TOCA1 HR1 domain also fully competed with the GST-ACK but bound with an affinity of 6 μm (Fig. 1, B and C), in agreement with the low affinity observed in the direct binding experiments.	RESULTS
+76	84	affinity	evidence	The TOCA1 HR1 domain also fully competed with the GST-ACK but bound with an affinity of 6 μm (Fig. 1, B and C), in agreement with the low affinity observed in the direct binding experiments.	RESULTS
+138	146	affinity	evidence	The TOCA1 HR1 domain also fully competed with the GST-ACK but bound with an affinity of 6 μm (Fig. 1, B and C), in agreement with the low affinity observed in the direct binding experiments.	RESULTS
+163	189	direct binding experiments	experimental_method	The TOCA1 HR1 domain also fully competed with the GST-ACK but bound with an affinity of 6 μm (Fig. 1, B and C), in agreement with the low affinity observed in the direct binding experiments.	RESULTS
+4	9	Cdc42	protein	The Cdc42 construct used in the binding assays has seven residues deleted from the C terminus to facilitate purification.	RESULTS
+32	46	binding assays	experimental_method	The Cdc42 construct used in the binding assays has seven residues deleted from the C terminus to facilitate purification.	RESULTS
+51	65	seven residues	residue_range	The Cdc42 construct used in the binding assays has seven residues deleted from the C terminus to facilitate purification.	RESULTS
+66	73	deleted	experimental_method	The Cdc42 construct used in the binding assays has seven residues deleted from the C terminus to facilitate purification.	RESULTS
+46	55	G protein	protein_type	These residues are not generally required for G protein-effector interactions, including the interaction between RhoA and the PRK1 HR1a domain.	RESULTS
+113	117	RhoA	protein	These residues are not generally required for G protein-effector interactions, including the interaction between RhoA and the PRK1 HR1a domain.	RESULTS
+126	130	PRK1	protein	These residues are not generally required for G protein-effector interactions, including the interaction between RhoA and the PRK1 HR1a domain.	RESULTS
+131	135	HR1a	structure_element	These residues are not generally required for G protein-effector interactions, including the interaction between RhoA and the PRK1 HR1a domain.	RESULTS
+31	35	Rac1	protein	In contrast, the C terminus of Rac1 contains a polybasic sequence, which is crucial for Rac1 binding to the HR1b domain from PRK1.	RESULTS
+88	92	Rac1	protein	In contrast, the C terminus of Rac1 contains a polybasic sequence, which is crucial for Rac1 binding to the HR1b domain from PRK1.	RESULTS
+108	112	HR1b	structure_element	In contrast, the C terminus of Rac1 contains a polybasic sequence, which is crucial for Rac1 binding to the HR1b domain from PRK1.	RESULTS
+125	129	PRK1	protein	In contrast, the C terminus of Rac1 contains a polybasic sequence, which is crucial for Rac1 binding to the HR1b domain from PRK1.	RESULTS
+16	24	affinity	evidence	As the observed affinity between TOCA1 HR1 and Cdc42 was much lower than expected, we reasoned that the C terminus of Cdc42 might be necessary for a high affinity interaction.	RESULTS
+33	38	TOCA1	protein	As the observed affinity between TOCA1 HR1 and Cdc42 was much lower than expected, we reasoned that the C terminus of Cdc42 might be necessary for a high affinity interaction.	RESULTS
+39	42	HR1	structure_element	As the observed affinity between TOCA1 HR1 and Cdc42 was much lower than expected, we reasoned that the C terminus of Cdc42 might be necessary for a high affinity interaction.	RESULTS
+47	52	Cdc42	protein	As the observed affinity between TOCA1 HR1 and Cdc42 was much lower than expected, we reasoned that the C terminus of Cdc42 might be necessary for a high affinity interaction.	RESULTS
+118	123	Cdc42	protein	As the observed affinity between TOCA1 HR1 and Cdc42 was much lower than expected, we reasoned that the C terminus of Cdc42 might be necessary for a high affinity interaction.	RESULTS
+154	162	affinity	evidence	As the observed affinity between TOCA1 HR1 and Cdc42 was much lower than expected, we reasoned that the C terminus of Cdc42 might be necessary for a high affinity interaction.	RESULTS
+4	23	binding experiments	experimental_method	The binding experiments were repeated with full-length [3H]GTP·Cdc42, but the affinity of the HR1 domain for full-length Cdc42 was similar to its affinity for truncated Cdc42 (Kd ≈ 5 μm; Fig. 1C).	RESULTS
+43	54	full-length	protein_state	The binding experiments were repeated with full-length [3H]GTP·Cdc42, but the affinity of the HR1 domain for full-length Cdc42 was similar to its affinity for truncated Cdc42 (Kd ≈ 5 μm; Fig. 1C).	RESULTS
+55	68	[3H]GTP·Cdc42	complex_assembly	The binding experiments were repeated with full-length [3H]GTP·Cdc42, but the affinity of the HR1 domain for full-length Cdc42 was similar to its affinity for truncated Cdc42 (Kd ≈ 5 μm; Fig. 1C).	RESULTS
+78	86	affinity	evidence	The binding experiments were repeated with full-length [3H]GTP·Cdc42, but the affinity of the HR1 domain for full-length Cdc42 was similar to its affinity for truncated Cdc42 (Kd ≈ 5 μm; Fig. 1C).	RESULTS
+94	97	HR1	structure_element	The binding experiments were repeated with full-length [3H]GTP·Cdc42, but the affinity of the HR1 domain for full-length Cdc42 was similar to its affinity for truncated Cdc42 (Kd ≈ 5 μm; Fig. 1C).	RESULTS
+109	120	full-length	protein_state	The binding experiments were repeated with full-length [3H]GTP·Cdc42, but the affinity of the HR1 domain for full-length Cdc42 was similar to its affinity for truncated Cdc42 (Kd ≈ 5 μm; Fig. 1C).	RESULTS
+121	126	Cdc42	protein	The binding experiments were repeated with full-length [3H]GTP·Cdc42, but the affinity of the HR1 domain for full-length Cdc42 was similar to its affinity for truncated Cdc42 (Kd ≈ 5 μm; Fig. 1C).	RESULTS
+146	154	affinity	evidence	The binding experiments were repeated with full-length [3H]GTP·Cdc42, but the affinity of the HR1 domain for full-length Cdc42 was similar to its affinity for truncated Cdc42 (Kd ≈ 5 μm; Fig. 1C).	RESULTS
+159	168	truncated	protein_state	The binding experiments were repeated with full-length [3H]GTP·Cdc42, but the affinity of the HR1 domain for full-length Cdc42 was similar to its affinity for truncated Cdc42 (Kd ≈ 5 μm; Fig. 1C).	RESULTS
+169	174	Cdc42	protein	The binding experiments were repeated with full-length [3H]GTP·Cdc42, but the affinity of the HR1 domain for full-length Cdc42 was similar to its affinity for truncated Cdc42 (Kd ≈ 5 μm; Fig. 1C).	RESULTS
+176	178	Kd	evidence	The binding experiments were repeated with full-length [3H]GTP·Cdc42, but the affinity of the HR1 domain for full-length Cdc42 was similar to its affinity for truncated Cdc42 (Kd ≈ 5 μm; Fig. 1C).	RESULTS
+10	27	C-terminal region	structure_element	Thus, the C-terminal region of Cdc42 is not required for maximal binding of TOCA1 HR1.	RESULTS
+31	36	Cdc42	protein	Thus, the C-terminal region of Cdc42 is not required for maximal binding of TOCA1 HR1.	RESULTS
+76	81	TOCA1	protein	Thus, the C-terminal region of Cdc42 is not required for maximal binding of TOCA1 HR1.	RESULTS
+82	85	HR1	structure_element	Thus, the C-terminal region of Cdc42 is not required for maximal binding of TOCA1 HR1.	RESULTS
+41	51	affinities	evidence	Another possible explanation for the low affinities observed was that the HR1 domain alone is not sufficient for maximal binding of the TOCA proteins to Cdc42 and that the other domains are required.	RESULTS
+74	77	HR1	structure_element	Another possible explanation for the low affinities observed was that the HR1 domain alone is not sufficient for maximal binding of the TOCA proteins to Cdc42 and that the other domains are required.	RESULTS
+85	90	alone	protein_state	Another possible explanation for the low affinities observed was that the HR1 domain alone is not sufficient for maximal binding of the TOCA proteins to Cdc42 and that the other domains are required.	RESULTS
+136	149	TOCA proteins	protein_type	Another possible explanation for the low affinities observed was that the HR1 domain alone is not sufficient for maximal binding of the TOCA proteins to Cdc42 and that the other domains are required.	RESULTS
+153	158	Cdc42	protein	Another possible explanation for the low affinities observed was that the HR1 domain alone is not sufficient for maximal binding of the TOCA proteins to Cdc42 and that the other domains are required.	RESULTS
+8	22	GST pull-downs	experimental_method	Indeed, GST pull-downs performed with in vitro translated human TOCA1 fragments had suggested that residues N-terminal to the HR1 domain may be required to stabilize the HR1 domain structure.	RESULTS
+58	63	human	species	Indeed, GST pull-downs performed with in vitro translated human TOCA1 fragments had suggested that residues N-terminal to the HR1 domain may be required to stabilize the HR1 domain structure.	RESULTS
+64	69	TOCA1	protein	Indeed, GST pull-downs performed with in vitro translated human TOCA1 fragments had suggested that residues N-terminal to the HR1 domain may be required to stabilize the HR1 domain structure.	RESULTS
+126	129	HR1	structure_element	Indeed, GST pull-downs performed with in vitro translated human TOCA1 fragments had suggested that residues N-terminal to the HR1 domain may be required to stabilize the HR1 domain structure.	RESULTS
+170	173	HR1	structure_element	Indeed, GST pull-downs performed with in vitro translated human TOCA1 fragments had suggested that residues N-terminal to the HR1 domain may be required to stabilize the HR1 domain structure.	RESULTS
+18	21	BAR	structure_element	Furthermore, both BAR and SH3 domains have been implicated in interactions with small G proteins (e.g. the BAR domain of Arfaptin2 binds to Rac1 and Arl1), while an SH3 domain mediates the interaction between Rac1 and the guanine nucleotide exchange factor, β-PIX.	RESULTS
+26	29	SH3	structure_element	Furthermore, both BAR and SH3 domains have been implicated in interactions with small G proteins (e.g. the BAR domain of Arfaptin2 binds to Rac1 and Arl1), while an SH3 domain mediates the interaction between Rac1 and the guanine nucleotide exchange factor, β-PIX.	RESULTS
+86	96	G proteins	protein_type	Furthermore, both BAR and SH3 domains have been implicated in interactions with small G proteins (e.g. the BAR domain of Arfaptin2 binds to Rac1 and Arl1), while an SH3 domain mediates the interaction between Rac1 and the guanine nucleotide exchange factor, β-PIX.	RESULTS
+107	110	BAR	structure_element	Furthermore, both BAR and SH3 domains have been implicated in interactions with small G proteins (e.g. the BAR domain of Arfaptin2 binds to Rac1 and Arl1), while an SH3 domain mediates the interaction between Rac1 and the guanine nucleotide exchange factor, β-PIX.	RESULTS
+121	130	Arfaptin2	protein	Furthermore, both BAR and SH3 domains have been implicated in interactions with small G proteins (e.g. the BAR domain of Arfaptin2 binds to Rac1 and Arl1), while an SH3 domain mediates the interaction between Rac1 and the guanine nucleotide exchange factor, β-PIX.	RESULTS
+140	144	Rac1	protein	Furthermore, both BAR and SH3 domains have been implicated in interactions with small G proteins (e.g. the BAR domain of Arfaptin2 binds to Rac1 and Arl1), while an SH3 domain mediates the interaction between Rac1 and the guanine nucleotide exchange factor, β-PIX.	RESULTS
+149	153	Arl1	protein	Furthermore, both BAR and SH3 domains have been implicated in interactions with small G proteins (e.g. the BAR domain of Arfaptin2 binds to Rac1 and Arl1), while an SH3 domain mediates the interaction between Rac1 and the guanine nucleotide exchange factor, β-PIX.	RESULTS
+165	168	SH3	structure_element	Furthermore, both BAR and SH3 domains have been implicated in interactions with small G proteins (e.g. the BAR domain of Arfaptin2 binds to Rac1 and Arl1), while an SH3 domain mediates the interaction between Rac1 and the guanine nucleotide exchange factor, β-PIX.	RESULTS
+209	213	Rac1	protein	Furthermore, both BAR and SH3 domains have been implicated in interactions with small G proteins (e.g. the BAR domain of Arfaptin2 binds to Rac1 and Arl1), while an SH3 domain mediates the interaction between Rac1 and the guanine nucleotide exchange factor, β-PIX.	RESULTS
+222	256	guanine nucleotide exchange factor	protein	Furthermore, both BAR and SH3 domains have been implicated in interactions with small G proteins (e.g. the BAR domain of Arfaptin2 binds to Rac1 and Arl1), while an SH3 domain mediates the interaction between Rac1 and the guanine nucleotide exchange factor, β-PIX.	RESULTS
+258	263	β-PIX	protein	Furthermore, both BAR and SH3 domains have been implicated in interactions with small G proteins (e.g. the BAR domain of Arfaptin2 binds to Rac1 and Arl1), while an SH3 domain mediates the interaction between Rac1 and the guanine nucleotide exchange factor, β-PIX.	RESULTS
+0	5	TOCA1	protein	TOCA1 dimerizes via its F-BAR domain, which could also affect Cdc42 binding, for example by presenting two HR1 domains for Cdc42 interactions.	RESULTS
+6	11	dimer	oligomeric_state	TOCA1 dimerizes via its F-BAR domain, which could also affect Cdc42 binding, for example by presenting two HR1 domains for Cdc42 interactions.	RESULTS
+24	29	F-BAR	structure_element	TOCA1 dimerizes via its F-BAR domain, which could also affect Cdc42 binding, for example by presenting two HR1 domains for Cdc42 interactions.	RESULTS
+62	67	Cdc42	protein	TOCA1 dimerizes via its F-BAR domain, which could also affect Cdc42 binding, for example by presenting two HR1 domains for Cdc42 interactions.	RESULTS
+107	110	HR1	structure_element	TOCA1 dimerizes via its F-BAR domain, which could also affect Cdc42 binding, for example by presenting two HR1 domains for Cdc42 interactions.	RESULTS
+123	128	Cdc42	protein	TOCA1 dimerizes via its F-BAR domain, which could also affect Cdc42 binding, for example by presenting two HR1 domains for Cdc42 interactions.	RESULTS
+8	13	TOCA1	protein	Various TOCA1 fragments (Fig. 2A) were therefore assessed for binding to full-length Cdc42 by direct SPA.	RESULTS
+73	84	full-length	protein_state	Various TOCA1 fragments (Fig. 2A) were therefore assessed for binding to full-length Cdc42 by direct SPA.	RESULTS
+85	90	Cdc42	protein	Various TOCA1 fragments (Fig. 2A) were therefore assessed for binding to full-length Cdc42 by direct SPA.	RESULTS
+101	104	SPA	experimental_method	Various TOCA1 fragments (Fig. 2A) were therefore assessed for binding to full-length Cdc42 by direct SPA.	RESULTS
+13	18	F-BAR	structure_element	The isolated F-BAR domain showed no binding to full-length Cdc42 (Fig. 2B).	RESULTS
+47	58	full-length	protein_state	The isolated F-BAR domain showed no binding to full-length Cdc42 (Fig. 2B).	RESULTS
+59	64	Cdc42	protein	The isolated F-BAR domain showed no binding to full-length Cdc42 (Fig. 2B).	RESULTS
+0	11	Full-length	protein_state	Full-length TOCA1 and ΔSH3 TOCA1 bound with micromolar affinity (Fig. 2B), in a similar manner to the isolated HR1 domain (Fig. 1A).	RESULTS
+12	17	TOCA1	protein	Full-length TOCA1 and ΔSH3 TOCA1 bound with micromolar affinity (Fig. 2B), in a similar manner to the isolated HR1 domain (Fig. 1A).	RESULTS
+22	26	ΔSH3	mutant	Full-length TOCA1 and ΔSH3 TOCA1 bound with micromolar affinity (Fig. 2B), in a similar manner to the isolated HR1 domain (Fig. 1A).	RESULTS
+27	32	TOCA1	protein	Full-length TOCA1 and ΔSH3 TOCA1 bound with micromolar affinity (Fig. 2B), in a similar manner to the isolated HR1 domain (Fig. 1A).	RESULTS
+33	38	bound	protein_state	Full-length TOCA1 and ΔSH3 TOCA1 bound with micromolar affinity (Fig. 2B), in a similar manner to the isolated HR1 domain (Fig. 1A).	RESULTS
+111	114	HR1	structure_element	Full-length TOCA1 and ΔSH3 TOCA1 bound with micromolar affinity (Fig. 2B), in a similar manner to the isolated HR1 domain (Fig. 1A).	RESULTS
+4	11	HR1-SH3	mutant	The HR1-SH3 protein could not be purified to homogeneity as a fusion protein, so it was assayed in competition assays after cleavage of the His tag.	RESULTS
+99	117	competition assays	experimental_method	The HR1-SH3 protein could not be purified to homogeneity as a fusion protein, so it was assayed in competition assays after cleavage of the His tag.	RESULTS
+29	36	GST-ACK	mutant	This construct competed with GST-ACK GBD to give a similar affinity to the HR1 domain alone (Kd = 4.6 ± 4 μm; Fig. 2C).	RESULTS
+37	40	GBD	structure_element	This construct competed with GST-ACK GBD to give a similar affinity to the HR1 domain alone (Kd = 4.6 ± 4 μm; Fig. 2C).	RESULTS
+75	78	HR1	structure_element	This construct competed with GST-ACK GBD to give a similar affinity to the HR1 domain alone (Kd = 4.6 ± 4 μm; Fig. 2C).	RESULTS
+86	91	alone	protein_state	This construct competed with GST-ACK GBD to give a similar affinity to the HR1 domain alone (Kd = 4.6 ± 4 μm; Fig. 2C).	RESULTS
+93	95	Kd	evidence	This construct competed with GST-ACK GBD to give a similar affinity to the HR1 domain alone (Kd = 4.6 ± 4 μm; Fig. 2C).	RESULTS
+44	49	TOCA1	protein	Taken together, these data suggest that the TOCA1 HR1 domain is sufficient for maximal binding and that this binding is of a relatively low affinity compared with many other Cdc42·effector complexes.	RESULTS
+50	53	HR1	structure_element	Taken together, these data suggest that the TOCA1 HR1 domain is sufficient for maximal binding and that this binding is of a relatively low affinity compared with many other Cdc42·effector complexes.	RESULTS
+174	179	Cdc42	protein	Taken together, these data suggest that the TOCA1 HR1 domain is sufficient for maximal binding and that this binding is of a relatively low affinity compared with many other Cdc42·effector complexes.	RESULTS
+4	13	Cdc42-HR1	complex_assembly	The Cdc42-HR1 interaction is of low affinity in the context of full-length protein and in TOCA1 paralogues.	FIG
+63	74	full-length	protein_state	The Cdc42-HR1 interaction is of low affinity in the context of full-length protein and in TOCA1 paralogues.	FIG
+90	95	TOCA1	protein	The Cdc42-HR1 interaction is of low affinity in the context of full-length protein and in TOCA1 paralogues.	FIG
+29	34	TOCA1	protein	 A, diagram illustrating the TOCA1 constructs assayed for Cdc42 binding.	FIG
+58	63	Cdc42	protein	 A, diagram illustrating the TOCA1 constructs assayed for Cdc42 binding.	FIG
+71	85	binding curves	evidence	Domain boundaries are derived from secondary structure predictions; B, binding curves derived from direct binding assays, in which the indicated concentrations of Cdc42Δ7Q61L·[3H]GTP were incubated with 30 nm GST-ACK or His-tagged TOCA1 constructs, as indicated, in SPAs.	FIG
+99	120	direct binding assays	experimental_method	Domain boundaries are derived from secondary structure predictions; B, binding curves derived from direct binding assays, in which the indicated concentrations of Cdc42Δ7Q61L·[3H]GTP were incubated with 30 nm GST-ACK or His-tagged TOCA1 constructs, as indicated, in SPAs.	FIG
+163	182	Cdc42Δ7Q61L·[3H]GTP	complex_assembly	Domain boundaries are derived from secondary structure predictions; B, binding curves derived from direct binding assays, in which the indicated concentrations of Cdc42Δ7Q61L·[3H]GTP were incubated with 30 nm GST-ACK or His-tagged TOCA1 constructs, as indicated, in SPAs.	FIG
+188	197	incubated	experimental_method	Domain boundaries are derived from secondary structure predictions; B, binding curves derived from direct binding assays, in which the indicated concentrations of Cdc42Δ7Q61L·[3H]GTP were incubated with 30 nm GST-ACK or His-tagged TOCA1 constructs, as indicated, in SPAs.	FIG
+209	216	GST-ACK	mutant	Domain boundaries are derived from secondary structure predictions; B, binding curves derived from direct binding assays, in which the indicated concentrations of Cdc42Δ7Q61L·[3H]GTP were incubated with 30 nm GST-ACK or His-tagged TOCA1 constructs, as indicated, in SPAs.	FIG
+220	230	His-tagged	protein_state	Domain boundaries are derived from secondary structure predictions; B, binding curves derived from direct binding assays, in which the indicated concentrations of Cdc42Δ7Q61L·[3H]GTP were incubated with 30 nm GST-ACK or His-tagged TOCA1 constructs, as indicated, in SPAs.	FIG
+231	236	TOCA1	protein	Domain boundaries are derived from secondary structure predictions; B, binding curves derived from direct binding assays, in which the indicated concentrations of Cdc42Δ7Q61L·[3H]GTP were incubated with 30 nm GST-ACK or His-tagged TOCA1 constructs, as indicated, in SPAs.	FIG
+266	270	SPAs	experimental_method	Domain boundaries are derived from secondary structure predictions; B, binding curves derived from direct binding assays, in which the indicated concentrations of Cdc42Δ7Q61L·[3H]GTP were incubated with 30 nm GST-ACK or His-tagged TOCA1 constructs, as indicated, in SPAs.	FIG
+4	7	SPA	experimental_method	The SPA signal was corrected by subtraction of control data with no fusion protein.	FIG
+26	42	binding isotherm	evidence	The data were fitted to a binding isotherm to give an apparent Kd and are expressed as a percentage of the maximum signal.	FIG
+63	65	Kd	evidence	The data were fitted to a binding isotherm to give an apparent Kd and are expressed as a percentage of the maximum signal.	FIG
+32	47	competition SPA	experimental_method	C–E, representative examples of competition SPA experiments carried out with the indicated concentrations of the TOCA1 HR1-SH3 construct titrated into 30 nm GST-ACK and 30 nm Cdc42Δ7Q61L·[3H]GTP (C) or HR1CIP4 (D) or HR1FBP17 (E) titrated into 30 nm GST-ACK and 30 nm Cdc42FLQ61L·[3H]GTP.	FIG
+113	118	TOCA1	protein	C–E, representative examples of competition SPA experiments carried out with the indicated concentrations of the TOCA1 HR1-SH3 construct titrated into 30 nm GST-ACK and 30 nm Cdc42Δ7Q61L·[3H]GTP (C) or HR1CIP4 (D) or HR1FBP17 (E) titrated into 30 nm GST-ACK and 30 nm Cdc42FLQ61L·[3H]GTP.	FIG
+119	126	HR1-SH3	mutant	C–E, representative examples of competition SPA experiments carried out with the indicated concentrations of the TOCA1 HR1-SH3 construct titrated into 30 nm GST-ACK and 30 nm Cdc42Δ7Q61L·[3H]GTP (C) or HR1CIP4 (D) or HR1FBP17 (E) titrated into 30 nm GST-ACK and 30 nm Cdc42FLQ61L·[3H]GTP.	FIG
+137	145	titrated	experimental_method	C–E, representative examples of competition SPA experiments carried out with the indicated concentrations of the TOCA1 HR1-SH3 construct titrated into 30 nm GST-ACK and 30 nm Cdc42Δ7Q61L·[3H]GTP (C) or HR1CIP4 (D) or HR1FBP17 (E) titrated into 30 nm GST-ACK and 30 nm Cdc42FLQ61L·[3H]GTP.	FIG
+157	164	GST-ACK	mutant	C–E, representative examples of competition SPA experiments carried out with the indicated concentrations of the TOCA1 HR1-SH3 construct titrated into 30 nm GST-ACK and 30 nm Cdc42Δ7Q61L·[3H]GTP (C) or HR1CIP4 (D) or HR1FBP17 (E) titrated into 30 nm GST-ACK and 30 nm Cdc42FLQ61L·[3H]GTP.	FIG
+175	194	Cdc42Δ7Q61L·[3H]GTP	complex_assembly	C–E, representative examples of competition SPA experiments carried out with the indicated concentrations of the TOCA1 HR1-SH3 construct titrated into 30 nm GST-ACK and 30 nm Cdc42Δ7Q61L·[3H]GTP (C) or HR1CIP4 (D) or HR1FBP17 (E) titrated into 30 nm GST-ACK and 30 nm Cdc42FLQ61L·[3H]GTP.	FIG
+202	205	HR1	structure_element	C–E, representative examples of competition SPA experiments carried out with the indicated concentrations of the TOCA1 HR1-SH3 construct titrated into 30 nm GST-ACK and 30 nm Cdc42Δ7Q61L·[3H]GTP (C) or HR1CIP4 (D) or HR1FBP17 (E) titrated into 30 nm GST-ACK and 30 nm Cdc42FLQ61L·[3H]GTP.	FIG
+217	220	HR1	structure_element	C–E, representative examples of competition SPA experiments carried out with the indicated concentrations of the TOCA1 HR1-SH3 construct titrated into 30 nm GST-ACK and 30 nm Cdc42Δ7Q61L·[3H]GTP (C) or HR1CIP4 (D) or HR1FBP17 (E) titrated into 30 nm GST-ACK and 30 nm Cdc42FLQ61L·[3H]GTP.	FIG
+230	238	titrated	experimental_method	C–E, representative examples of competition SPA experiments carried out with the indicated concentrations of the TOCA1 HR1-SH3 construct titrated into 30 nm GST-ACK and 30 nm Cdc42Δ7Q61L·[3H]GTP (C) or HR1CIP4 (D) or HR1FBP17 (E) titrated into 30 nm GST-ACK and 30 nm Cdc42FLQ61L·[3H]GTP.	FIG
+250	257	GST-ACK	mutant	C–E, representative examples of competition SPA experiments carried out with the indicated concentrations of the TOCA1 HR1-SH3 construct titrated into 30 nm GST-ACK and 30 nm Cdc42Δ7Q61L·[3H]GTP (C) or HR1CIP4 (D) or HR1FBP17 (E) titrated into 30 nm GST-ACK and 30 nm Cdc42FLQ61L·[3H]GTP.	FIG
+268	287	Cdc42FLQ61L·[3H]GTP	complex_assembly	C–E, representative examples of competition SPA experiments carried out with the indicated concentrations of the TOCA1 HR1-SH3 construct titrated into 30 nm GST-ACK and 30 nm Cdc42Δ7Q61L·[3H]GTP (C) or HR1CIP4 (D) or HR1FBP17 (E) titrated into 30 nm GST-ACK and 30 nm Cdc42FLQ61L·[3H]GTP.	FIG
+24	29	TOCA1	protein	The low affinity of the TOCA1 HR1-Cdc42 interaction raised the question of whether the other known Cdc42-binding TOCA family proteins, FBP17 and CIP4, also bind weakly.	RESULTS
+30	33	HR1	structure_element	The low affinity of the TOCA1 HR1-Cdc42 interaction raised the question of whether the other known Cdc42-binding TOCA family proteins, FBP17 and CIP4, also bind weakly.	RESULTS
+34	39	Cdc42	protein	The low affinity of the TOCA1 HR1-Cdc42 interaction raised the question of whether the other known Cdc42-binding TOCA family proteins, FBP17 and CIP4, also bind weakly.	RESULTS
+99	104	Cdc42	protein	The low affinity of the TOCA1 HR1-Cdc42 interaction raised the question of whether the other known Cdc42-binding TOCA family proteins, FBP17 and CIP4, also bind weakly.	RESULTS
+113	133	TOCA family proteins	protein_type	The low affinity of the TOCA1 HR1-Cdc42 interaction raised the question of whether the other known Cdc42-binding TOCA family proteins, FBP17 and CIP4, also bind weakly.	RESULTS
+135	140	FBP17	protein	The low affinity of the TOCA1 HR1-Cdc42 interaction raised the question of whether the other known Cdc42-binding TOCA family proteins, FBP17 and CIP4, also bind weakly.	RESULTS
+145	149	CIP4	protein	The low affinity of the TOCA1 HR1-Cdc42 interaction raised the question of whether the other known Cdc42-binding TOCA family proteins, FBP17 and CIP4, also bind weakly.	RESULTS
+4	7	HR1	structure_element	The HR1 domains from FBP17 and CIP4 were purified and assayed for Cdc42 binding in competition SPAs, analogous to those carried out with the TOCA1 HR1 domain.	RESULTS
+21	26	FBP17	protein	The HR1 domains from FBP17 and CIP4 were purified and assayed for Cdc42 binding in competition SPAs, analogous to those carried out with the TOCA1 HR1 domain.	RESULTS
+31	35	CIP4	protein	The HR1 domains from FBP17 and CIP4 were purified and assayed for Cdc42 binding in competition SPAs, analogous to those carried out with the TOCA1 HR1 domain.	RESULTS
+41	49	purified	experimental_method	The HR1 domains from FBP17 and CIP4 were purified and assayed for Cdc42 binding in competition SPAs, analogous to those carried out with the TOCA1 HR1 domain.	RESULTS
+66	71	Cdc42	protein	The HR1 domains from FBP17 and CIP4 were purified and assayed for Cdc42 binding in competition SPAs, analogous to those carried out with the TOCA1 HR1 domain.	RESULTS
+83	99	competition SPAs	experimental_method	The HR1 domains from FBP17 and CIP4 were purified and assayed for Cdc42 binding in competition SPAs, analogous to those carried out with the TOCA1 HR1 domain.	RESULTS
+141	146	TOCA1	protein	The HR1 domains from FBP17 and CIP4 were purified and assayed for Cdc42 binding in competition SPAs, analogous to those carried out with the TOCA1 HR1 domain.	RESULTS
+147	150	HR1	structure_element	The HR1 domains from FBP17 and CIP4 were purified and assayed for Cdc42 binding in competition SPAs, analogous to those carried out with the TOCA1 HR1 domain.	RESULTS
+4	14	affinities	evidence	The affinities of both the FBP17 and CIP4 HR1 domains were also in the low micromolar range (10 and 5 μm, respectively) (Fig. 2, D and E), suggesting that low affinity interactions with Cdc42 are a common feature within the TOCA family.	RESULTS
+27	32	FBP17	protein	The affinities of both the FBP17 and CIP4 HR1 domains were also in the low micromolar range (10 and 5 μm, respectively) (Fig. 2, D and E), suggesting that low affinity interactions with Cdc42 are a common feature within the TOCA family.	RESULTS
+37	41	CIP4	protein	The affinities of both the FBP17 and CIP4 HR1 domains were also in the low micromolar range (10 and 5 μm, respectively) (Fig. 2, D and E), suggesting that low affinity interactions with Cdc42 are a common feature within the TOCA family.	RESULTS
+42	45	HR1	structure_element	The affinities of both the FBP17 and CIP4 HR1 domains were also in the low micromolar range (10 and 5 μm, respectively) (Fig. 2, D and E), suggesting that low affinity interactions with Cdc42 are a common feature within the TOCA family.	RESULTS
+186	191	Cdc42	protein	The affinities of both the FBP17 and CIP4 HR1 domains were also in the low micromolar range (10 and 5 μm, respectively) (Fig. 2, D and E), suggesting that low affinity interactions with Cdc42 are a common feature within the TOCA family.	RESULTS
+224	235	TOCA family	protein_type	The affinities of both the FBP17 and CIP4 HR1 domains were also in the low micromolar range (10 and 5 μm, respectively) (Fig. 2, D and E), suggesting that low affinity interactions with Cdc42 are a common feature within the TOCA family.	RESULTS
+0	9	Structure	evidence	Structure of the TOCA1 HR1 Domain	RESULTS
+17	22	TOCA1	protein	Structure of the TOCA1 HR1 Domain	RESULTS
+23	26	HR1	structure_element	Structure of the TOCA1 HR1 Domain	RESULTS
+12	17	TOCA1	protein	Because the TOCA1 HR1 domain was sufficient for maximal Cdc42-binding, we used this construct for structural studies.	RESULTS
+18	21	HR1	structure_element	Because the TOCA1 HR1 domain was sufficient for maximal Cdc42-binding, we used this construct for structural studies.	RESULTS
+56	61	Cdc42	protein	Because the TOCA1 HR1 domain was sufficient for maximal Cdc42-binding, we used this construct for structural studies.	RESULTS
+40	45	TOCA1	protein	Initial experiments were performed with TOCA1 residues 324–426, but we observed that the N terminus was cleaved during purification to yield a new N terminus at residue 330 (data not shown).	RESULTS
+55	62	324–426	residue_range	Initial experiments were performed with TOCA1 residues 324–426, but we observed that the N terminus was cleaved during purification to yield a new N terminus at residue 330 (data not shown).	RESULTS
+169	172	330	residue_number	Initial experiments were performed with TOCA1 residues 324–426, but we observed that the N terminus was cleaved during purification to yield a new N terminus at residue 330 (data not shown).	RESULTS
+56	63	330–426	residue_range	We therefore engineered a construct comprising residues 330–426 to produce the minimal, stable HR1 domain.	RESULTS
+79	86	minimal	protein_state	We therefore engineered a construct comprising residues 330–426 to produce the minimal, stable HR1 domain.	RESULTS
+88	94	stable	protein_state	We therefore engineered a construct comprising residues 330–426 to produce the minimal, stable HR1 domain.	RESULTS
+95	98	HR1	structure_element	We therefore engineered a construct comprising residues 330–426 to produce the minimal, stable HR1 domain.	RESULTS
+21	35	NOE restraints	evidence	2,778 non-degenerate NOE restraints were used in initial structure calculations (1,791 unambiguous and 987 ambiguous), derived from three-dimensional 15N-separated NOESY and 13C-separated NOESY experiments.	RESULTS
+57	79	structure calculations	experimental_method	2,778 non-degenerate NOE restraints were used in initial structure calculations (1,791 unambiguous and 987 ambiguous), derived from three-dimensional 15N-separated NOESY and 13C-separated NOESY experiments.	RESULTS
+150	169	15N-separated NOESY	experimental_method	2,778 non-degenerate NOE restraints were used in initial structure calculations (1,791 unambiguous and 987 ambiguous), derived from three-dimensional 15N-separated NOESY and 13C-separated NOESY experiments.	RESULTS
+174	193	13C-separated NOESY	experimental_method	2,778 non-degenerate NOE restraints were used in initial structure calculations (1,791 unambiguous and 987 ambiguous), derived from three-dimensional 15N-separated NOESY and 13C-separated NOESY experiments.	RESULTS
+29	33	NOEs	evidence	There were 1,845 unambiguous NOEs and 757 ambiguous NOEs after eight iterations.	RESULTS
+52	56	NOEs	evidence	There were 1,845 unambiguous NOEs and 757 ambiguous NOEs after eight iterations.	RESULTS
+4	14	structures	evidence	100 structures were calculated in the final iteration; the 50 lowest energy structures were water-refined; and of these, the 35 lowest energy structures were analyzed.	RESULTS
+20	30	calculated	experimental_method	100 structures were calculated in the final iteration; the 50 lowest energy structures were water-refined; and of these, the 35 lowest energy structures were analyzed.	RESULTS
+76	86	structures	evidence	100 structures were calculated in the final iteration; the 50 lowest energy structures were water-refined; and of these, the 35 lowest energy structures were analyzed.	RESULTS
+142	152	structures	evidence	100 structures were calculated in the final iteration; the 50 lowest energy structures were water-refined; and of these, the 35 lowest energy structures were analyzed.	RESULTS
+27	30	HR1	structure_element	Table 1 indicates that the HR1 domain structure is well defined by the NMR data.	RESULTS
+38	47	structure	evidence	Table 1 indicates that the HR1 domain structure is well defined by the NMR data.	RESULTS
+71	74	NMR	experimental_method	Table 1 indicates that the HR1 domain structure is well defined by the NMR data.	RESULTS
+12	47	average root mean square deviations	evidence	a <SA>, the average root mean square deviations for the ensemble ± S.D.	TABLE
+24	33	structure	evidence	b <SA>c, values for the structure that is closest to the mean.	TABLE
+4	13	structure	evidence	The structure closest to the mean is shown in Fig. 3A.	RESULTS
+8	17	α-helices	structure_element	The two α-helices of the HR1 domain interact to form an anti-parallel coiled-coil with a slight left-handed twist, reminiscent of the HR1 domains of CIP4 (PDB code 2KE4) and PRK1 (PDB codes 1CXZ and 1URF).	RESULTS
+25	28	HR1	structure_element	The two α-helices of the HR1 domain interact to form an anti-parallel coiled-coil with a slight left-handed twist, reminiscent of the HR1 domains of CIP4 (PDB code 2KE4) and PRK1 (PDB codes 1CXZ and 1URF).	RESULTS
+56	81	anti-parallel coiled-coil	structure_element	The two α-helices of the HR1 domain interact to form an anti-parallel coiled-coil with a slight left-handed twist, reminiscent of the HR1 domains of CIP4 (PDB code 2KE4) and PRK1 (PDB codes 1CXZ and 1URF).	RESULTS
+134	137	HR1	structure_element	The two α-helices of the HR1 domain interact to form an anti-parallel coiled-coil with a slight left-handed twist, reminiscent of the HR1 domains of CIP4 (PDB code 2KE4) and PRK1 (PDB codes 1CXZ and 1URF).	RESULTS
+149	153	CIP4	protein	The two α-helices of the HR1 domain interact to form an anti-parallel coiled-coil with a slight left-handed twist, reminiscent of the HR1 domains of CIP4 (PDB code 2KE4) and PRK1 (PDB codes 1CXZ and 1URF).	RESULTS
+174	178	PRK1	protein	The two α-helices of the HR1 domain interact to form an anti-parallel coiled-coil with a slight left-handed twist, reminiscent of the HR1 domains of CIP4 (PDB code 2KE4) and PRK1 (PDB codes 1CXZ and 1URF).	RESULTS
+2	20	sequence alignment	experimental_method	A sequence alignment illustrating the secondary structure elements of the TOCA1 and CIP4 HR1 domains and the HR1a and HR1b domains from PRK1 is shown in Fig. 3B.	RESULTS
+74	79	TOCA1	protein	A sequence alignment illustrating the secondary structure elements of the TOCA1 and CIP4 HR1 domains and the HR1a and HR1b domains from PRK1 is shown in Fig. 3B.	RESULTS
+84	88	CIP4	protein	A sequence alignment illustrating the secondary structure elements of the TOCA1 and CIP4 HR1 domains and the HR1a and HR1b domains from PRK1 is shown in Fig. 3B.	RESULTS
+89	92	HR1	structure_element	A sequence alignment illustrating the secondary structure elements of the TOCA1 and CIP4 HR1 domains and the HR1a and HR1b domains from PRK1 is shown in Fig. 3B.	RESULTS
+109	113	HR1a	structure_element	A sequence alignment illustrating the secondary structure elements of the TOCA1 and CIP4 HR1 domains and the HR1a and HR1b domains from PRK1 is shown in Fig. 3B.	RESULTS
+118	122	HR1b	structure_element	A sequence alignment illustrating the secondary structure elements of the TOCA1 and CIP4 HR1 domains and the HR1a and HR1b domains from PRK1 is shown in Fig. 3B.	RESULTS
+136	140	PRK1	protein	A sequence alignment illustrating the secondary structure elements of the TOCA1 and CIP4 HR1 domains and the HR1a and HR1b domains from PRK1 is shown in Fig. 3B.	RESULTS
+4	13	structure	evidence	The structure of the TOCA1 HR1 domain.	FIG
+21	26	TOCA1	protein	The structure of the TOCA1 HR1 domain.	FIG
+27	30	HR1	structure_element	The structure of the TOCA1 HR1 domain.	FIG
+17	22	trace	evidence	 A, the backbone trace of the 35 lowest energy structures of the HR1 domain overlaid with the structure closest to the mean is shown alongside a schematic representation of the structure closest to the mean.	FIG
+47	57	structures	evidence	 A, the backbone trace of the 35 lowest energy structures of the HR1 domain overlaid with the structure closest to the mean is shown alongside a schematic representation of the structure closest to the mean.	FIG
+65	68	HR1	structure_element	 A, the backbone trace of the 35 lowest energy structures of the HR1 domain overlaid with the structure closest to the mean is shown alongside a schematic representation of the structure closest to the mean.	FIG
+94	103	structure	evidence	 A, the backbone trace of the 35 lowest energy structures of the HR1 domain overlaid with the structure closest to the mean is shown alongside a schematic representation of the structure closest to the mean.	FIG
+177	186	structure	evidence	 A, the backbone trace of the 35 lowest energy structures of the HR1 domain overlaid with the structure closest to the mean is shown alongside a schematic representation of the structure closest to the mean.	FIG
+50	57	330–333	residue_range	Flexible regions at the N and C termini (residues 330–333 and 421–426) are omitted for clarity.	FIG
+62	69	421–426	residue_range	Flexible regions at the N and C termini (residues 330–333 and 421–426) are omitted for clarity.	FIG
+5	23	sequence alignment	experimental_method	B, a sequence alignment of the HR1 domains from TOCA1, CIP4, and PRK1.	FIG
+31	34	HR1	structure_element	B, a sequence alignment of the HR1 domains from TOCA1, CIP4, and PRK1.	FIG
+48	53	TOCA1	protein	B, a sequence alignment of the HR1 domains from TOCA1, CIP4, and PRK1.	FIG
+55	59	CIP4	protein	B, a sequence alignment of the HR1 domains from TOCA1, CIP4, and PRK1.	FIG
+65	69	PRK1	protein	B, a sequence alignment of the HR1 domains from TOCA1, CIP4, and PRK1.	FIG
+42	48	Stride	experimental_method	The secondary structure was deduced using Stride, based on the Ramachandran angles, and is indicated as follows: gray, turn; yellow, α-helix; blue, 310 helix; white, coil.	FIG
+63	82	Ramachandran angles	evidence	The secondary structure was deduced using Stride, based on the Ramachandran angles, and is indicated as follows: gray, turn; yellow, α-helix; blue, 310 helix; white, coil.	FIG
+133	140	α-helix	structure_element	The secondary structure was deduced using Stride, based on the Ramachandran angles, and is indicated as follows: gray, turn; yellow, α-helix; blue, 310 helix; white, coil.	FIG
+148	157	310 helix	structure_element	The secondary structure was deduced using Stride, based on the Ramachandran angles, and is indicated as follows: gray, turn; yellow, α-helix; blue, 310 helix; white, coil.	FIG
+42	47	TOCA1	protein	C, a close-up of the N-terminal region of TOCA1 HR1, indicating some of the NOEs defining its position with respect to the two α-helices.	FIG
+48	51	HR1	structure_element	C, a close-up of the N-terminal region of TOCA1 HR1, indicating some of the NOEs defining its position with respect to the two α-helices.	FIG
+76	80	NOEs	evidence	C, a close-up of the N-terminal region of TOCA1 HR1, indicating some of the NOEs defining its position with respect to the two α-helices.	FIG
+127	136	α-helices	structure_element	C, a close-up of the N-terminal region of TOCA1 HR1, indicating some of the NOEs defining its position with respect to the two α-helices.	FIG
+14	28	NOE restraints	evidence	Dotted lines, NOE restraints.	FIG
+21	36	interhelix loop	structure_element	D, a close-up of the interhelix loop region showing some of the contacts between the loop and helix 1.	FIG
+85	89	loop	structure_element	D, a close-up of the interhelix loop region showing some of the contacts between the loop and helix 1.	FIG
+94	101	helix 1	structure_element	D, a close-up of the interhelix loop region showing some of the contacts between the loop and helix 1.	FIG
+7	11	HR1a	structure_element	In the HR1a domain of PRK1, a region N-terminal to helix 1 forms a short α-helix, which packs against both helices of the HR1 domain.	RESULTS
+22	26	PRK1	protein	In the HR1a domain of PRK1, a region N-terminal to helix 1 forms a short α-helix, which packs against both helices of the HR1 domain.	RESULTS
+51	58	helix 1	structure_element	In the HR1a domain of PRK1, a region N-terminal to helix 1 forms a short α-helix, which packs against both helices of the HR1 domain.	RESULTS
+67	80	short α-helix	structure_element	In the HR1a domain of PRK1, a region N-terminal to helix 1 forms a short α-helix, which packs against both helices of the HR1 domain.	RESULTS
+122	125	HR1	structure_element	In the HR1a domain of PRK1, a region N-terminal to helix 1 forms a short α-helix, which packs against both helices of the HR1 domain.	RESULTS
+15	20	TOCA1	protein	This region of TOCA1 HR1 (residues 334–340) is well defined in the family of structures (Fig. 3A) but does not form an α-helix.	RESULTS
+21	24	HR1	structure_element	This region of TOCA1 HR1 (residues 334–340) is well defined in the family of structures (Fig. 3A) but does not form an α-helix.	RESULTS
+35	42	334–340	residue_range	This region of TOCA1 HR1 (residues 334–340) is well defined in the family of structures (Fig. 3A) but does not form an α-helix.	RESULTS
+77	87	structures	evidence	This region of TOCA1 HR1 (residues 334–340) is well defined in the family of structures (Fig. 3A) but does not form an α-helix.	RESULTS
+119	126	α-helix	structure_element	This region of TOCA1 HR1 (residues 334–340) is well defined in the family of structures (Fig. 3A) but does not form an α-helix.	RESULTS
+47	61	NOE restraints	evidence	It instead forms a series of turns, defined by NOE restraints observed between residues separated by one (residues 332–334, 333–335, etc.) or two (residues 337–340) residues in the sequence and the φ and ψ angles, assessed using Stride.	RESULTS
+115	122	332–334	residue_range	It instead forms a series of turns, defined by NOE restraints observed between residues separated by one (residues 332–334, 333–335, etc.) or two (residues 337–340) residues in the sequence and the φ and ψ angles, assessed using Stride.	RESULTS
+124	131	333–335	residue_range	It instead forms a series of turns, defined by NOE restraints observed between residues separated by one (residues 332–334, 333–335, etc.) or two (residues 337–340) residues in the sequence and the φ and ψ angles, assessed using Stride.	RESULTS
+156	163	337–340	residue_range	It instead forms a series of turns, defined by NOE restraints observed between residues separated by one (residues 332–334, 333–335, etc.) or two (residues 337–340) residues in the sequence and the φ and ψ angles, assessed using Stride.	RESULTS
+198	212	φ and ψ angles	evidence	It instead forms a series of turns, defined by NOE restraints observed between residues separated by one (residues 332–334, 333–335, etc.) or two (residues 337–340) residues in the sequence and the φ and ψ angles, assessed using Stride.	RESULTS
+229	235	Stride	experimental_method	It instead forms a series of turns, defined by NOE restraints observed between residues separated by one (residues 332–334, 333–335, etc.) or two (residues 337–340) residues in the sequence and the φ and ψ angles, assessed using Stride.	RESULTS
+92	99	334–340	residue_range	These turns cause the chain to reverse direction, allowing the N-terminal segment (residues 334–340) to contact both helices of the HR1 domain.	RESULTS
+132	135	HR1	structure_element	These turns cause the chain to reverse direction, allowing the N-terminal segment (residues 334–340) to contact both helices of the HR1 domain.	RESULTS
+11	15	NOEs	evidence	Long range NOEs were observed linking Leu-334, Glu-335, and Asp-336 with Trp-413 of helix 2, Leu-334 with Lys-409 of helix 2, and Phe-337 and Ser-338 with Arg-345, Arg-348, and Leu-349 of helix 1.	RESULTS
+38	45	Leu-334	residue_name_number	Long range NOEs were observed linking Leu-334, Glu-335, and Asp-336 with Trp-413 of helix 2, Leu-334 with Lys-409 of helix 2, and Phe-337 and Ser-338 with Arg-345, Arg-348, and Leu-349 of helix 1.	RESULTS
+47	54	Glu-335	residue_name_number	Long range NOEs were observed linking Leu-334, Glu-335, and Asp-336 with Trp-413 of helix 2, Leu-334 with Lys-409 of helix 2, and Phe-337 and Ser-338 with Arg-345, Arg-348, and Leu-349 of helix 1.	RESULTS
+60	67	Asp-336	residue_name_number	Long range NOEs were observed linking Leu-334, Glu-335, and Asp-336 with Trp-413 of helix 2, Leu-334 with Lys-409 of helix 2, and Phe-337 and Ser-338 with Arg-345, Arg-348, and Leu-349 of helix 1.	RESULTS
+73	80	Trp-413	residue_name_number	Long range NOEs were observed linking Leu-334, Glu-335, and Asp-336 with Trp-413 of helix 2, Leu-334 with Lys-409 of helix 2, and Phe-337 and Ser-338 with Arg-345, Arg-348, and Leu-349 of helix 1.	RESULTS
+84	91	helix 2	structure_element	Long range NOEs were observed linking Leu-334, Glu-335, and Asp-336 with Trp-413 of helix 2, Leu-334 with Lys-409 of helix 2, and Phe-337 and Ser-338 with Arg-345, Arg-348, and Leu-349 of helix 1.	RESULTS
+93	100	Leu-334	residue_name_number	Long range NOEs were observed linking Leu-334, Glu-335, and Asp-336 with Trp-413 of helix 2, Leu-334 with Lys-409 of helix 2, and Phe-337 and Ser-338 with Arg-345, Arg-348, and Leu-349 of helix 1.	RESULTS
+106	113	Lys-409	residue_name_number	Long range NOEs were observed linking Leu-334, Glu-335, and Asp-336 with Trp-413 of helix 2, Leu-334 with Lys-409 of helix 2, and Phe-337 and Ser-338 with Arg-345, Arg-348, and Leu-349 of helix 1.	RESULTS
+117	124	helix 2	structure_element	Long range NOEs were observed linking Leu-334, Glu-335, and Asp-336 with Trp-413 of helix 2, Leu-334 with Lys-409 of helix 2, and Phe-337 and Ser-338 with Arg-345, Arg-348, and Leu-349 of helix 1.	RESULTS
+130	137	Phe-337	residue_name_number	Long range NOEs were observed linking Leu-334, Glu-335, and Asp-336 with Trp-413 of helix 2, Leu-334 with Lys-409 of helix 2, and Phe-337 and Ser-338 with Arg-345, Arg-348, and Leu-349 of helix 1.	RESULTS
+142	149	Ser-338	residue_name_number	Long range NOEs were observed linking Leu-334, Glu-335, and Asp-336 with Trp-413 of helix 2, Leu-334 with Lys-409 of helix 2, and Phe-337 and Ser-338 with Arg-345, Arg-348, and Leu-349 of helix 1.	RESULTS
+155	162	Arg-345	residue_name_number	Long range NOEs were observed linking Leu-334, Glu-335, and Asp-336 with Trp-413 of helix 2, Leu-334 with Lys-409 of helix 2, and Phe-337 and Ser-338 with Arg-345, Arg-348, and Leu-349 of helix 1.	RESULTS
+164	171	Arg-348	residue_name_number	Long range NOEs were observed linking Leu-334, Glu-335, and Asp-336 with Trp-413 of helix 2, Leu-334 with Lys-409 of helix 2, and Phe-337 and Ser-338 with Arg-345, Arg-348, and Leu-349 of helix 1.	RESULTS
+177	184	Leu-349	residue_name_number	Long range NOEs were observed linking Leu-334, Glu-335, and Asp-336 with Trp-413 of helix 2, Leu-334 with Lys-409 of helix 2, and Phe-337 and Ser-338 with Arg-345, Arg-348, and Leu-349 of helix 1.	RESULTS
+188	195	helix 1	structure_element	Long range NOEs were observed linking Leu-334, Glu-335, and Asp-336 with Trp-413 of helix 2, Leu-334 with Lys-409 of helix 2, and Phe-337 and Ser-338 with Arg-345, Arg-348, and Leu-349 of helix 1.	RESULTS
+8	17	α-helices	structure_element	The two α-helices of TOCA1 HR1 are separated by a long loop of 10 residues (residues 380–389) that contains two short 310 helices (residues 381–383 and 386–389).	RESULTS
+21	26	TOCA1	protein	The two α-helices of TOCA1 HR1 are separated by a long loop of 10 residues (residues 380–389) that contains two short 310 helices (residues 381–383 and 386–389).	RESULTS
+27	30	HR1	structure_element	The two α-helices of TOCA1 HR1 are separated by a long loop of 10 residues (residues 380–389) that contains two short 310 helices (residues 381–383 and 386–389).	RESULTS
+55	59	loop	structure_element	The two α-helices of TOCA1 HR1 are separated by a long loop of 10 residues (residues 380–389) that contains two short 310 helices (residues 381–383 and 386–389).	RESULTS
+85	92	380–389	residue_range	The two α-helices of TOCA1 HR1 are separated by a long loop of 10 residues (residues 380–389) that contains two short 310 helices (residues 381–383 and 386–389).	RESULTS
+112	129	short 310 helices	structure_element	The two α-helices of TOCA1 HR1 are separated by a long loop of 10 residues (residues 380–389) that contains two short 310 helices (residues 381–383 and 386–389).	RESULTS
+140	147	381–383	residue_range	The two α-helices of TOCA1 HR1 are separated by a long loop of 10 residues (residues 380–389) that contains two short 310 helices (residues 381–383 and 386–389).	RESULTS
+152	159	386–389	residue_range	The two α-helices of TOCA1 HR1 are separated by a long loop of 10 residues (residues 380–389) that contains two short 310 helices (residues 381–383 and 386–389).	RESULTS
+50	61	loop region	structure_element	Interestingly, side chains of residues within the loop region point back toward helix 1; for example, there are numerous distinct NOEs between the side chains of Asn-380 and Met-383 of the loop region and Tyr-377 and Val-376 of helix 1 (Fig. 3D).	RESULTS
+80	87	helix 1	structure_element	Interestingly, side chains of residues within the loop region point back toward helix 1; for example, there are numerous distinct NOEs between the side chains of Asn-380 and Met-383 of the loop region and Tyr-377 and Val-376 of helix 1 (Fig. 3D).	RESULTS
+162	169	Asn-380	residue_name_number	Interestingly, side chains of residues within the loop region point back toward helix 1; for example, there are numerous distinct NOEs between the side chains of Asn-380 and Met-383 of the loop region and Tyr-377 and Val-376 of helix 1 (Fig. 3D).	RESULTS
+174	181	Met-383	residue_name_number	Interestingly, side chains of residues within the loop region point back toward helix 1; for example, there are numerous distinct NOEs between the side chains of Asn-380 and Met-383 of the loop region and Tyr-377 and Val-376 of helix 1 (Fig. 3D).	RESULTS
+189	200	loop region	structure_element	Interestingly, side chains of residues within the loop region point back toward helix 1; for example, there are numerous distinct NOEs between the side chains of Asn-380 and Met-383 of the loop region and Tyr-377 and Val-376 of helix 1 (Fig. 3D).	RESULTS
+205	212	Tyr-377	residue_name_number	Interestingly, side chains of residues within the loop region point back toward helix 1; for example, there are numerous distinct NOEs between the side chains of Asn-380 and Met-383 of the loop region and Tyr-377 and Val-376 of helix 1 (Fig. 3D).	RESULTS
+217	224	Val-376	residue_name_number	Interestingly, side chains of residues within the loop region point back toward helix 1; for example, there are numerous distinct NOEs between the side chains of Asn-380 and Met-383 of the loop region and Tyr-377 and Val-376 of helix 1 (Fig. 3D).	RESULTS
+228	235	helix 1	structure_element	Interestingly, side chains of residues within the loop region point back toward helix 1; for example, there are numerous distinct NOEs between the side chains of Asn-380 and Met-383 of the loop region and Tyr-377 and Val-376 of helix 1 (Fig. 3D).	RESULTS
+34	41	Gly-384	residue_name_number	The backbone NH and CHα groups of Gly-384 and Asp-385 also show NOEs with the side chain of Tyr-377.	RESULTS
+46	53	Asp-385	residue_name_number	The backbone NH and CHα groups of Gly-384 and Asp-385 also show NOEs with the side chain of Tyr-377.	RESULTS
+92	99	Tyr-377	residue_name_number	The backbone NH and CHα groups of Gly-384 and Asp-385 also show NOEs with the side chain of Tyr-377.	RESULTS
+12	17	TOCA1	protein	Mapping the TOCA1 and Cdc42 Binding Interfaces	RESULTS
+22	46	Cdc42 Binding Interfaces	site	Mapping the TOCA1 and Cdc42 Binding Interfaces	RESULTS
+4	28	HR1TOCA1-Cdc42 interface	site	The HR1TOCA1-Cdc42 interface was investigated using NMR spectroscopy.	RESULTS
+52	68	NMR spectroscopy	experimental_method	The HR1TOCA1-Cdc42 interface was investigated using NMR spectroscopy.	RESULTS
+12	20	15N HSQC	experimental_method	A series of 15N HSQC experiments was recorded on 15N-labeled TOCA1 HR1 domain in the presence of increasing concentrations of unlabeled Cdc42Δ7Q61L·GMPPNP to map the Cdc42-binding surface.	RESULTS
+49	52	15N	chemical	A series of 15N HSQC experiments was recorded on 15N-labeled TOCA1 HR1 domain in the presence of increasing concentrations of unlabeled Cdc42Δ7Q61L·GMPPNP to map the Cdc42-binding surface.	RESULTS
+53	60	labeled	protein_state	A series of 15N HSQC experiments was recorded on 15N-labeled TOCA1 HR1 domain in the presence of increasing concentrations of unlabeled Cdc42Δ7Q61L·GMPPNP to map the Cdc42-binding surface.	RESULTS
+61	66	TOCA1	protein	A series of 15N HSQC experiments was recorded on 15N-labeled TOCA1 HR1 domain in the presence of increasing concentrations of unlabeled Cdc42Δ7Q61L·GMPPNP to map the Cdc42-binding surface.	RESULTS
+67	70	HR1	structure_element	A series of 15N HSQC experiments was recorded on 15N-labeled TOCA1 HR1 domain in the presence of increasing concentrations of unlabeled Cdc42Δ7Q61L·GMPPNP to map the Cdc42-binding surface.	RESULTS
+85	96	presence of	protein_state	A series of 15N HSQC experiments was recorded on 15N-labeled TOCA1 HR1 domain in the presence of increasing concentrations of unlabeled Cdc42Δ7Q61L·GMPPNP to map the Cdc42-binding surface.	RESULTS
+97	122	increasing concentrations	experimental_method	A series of 15N HSQC experiments was recorded on 15N-labeled TOCA1 HR1 domain in the presence of increasing concentrations of unlabeled Cdc42Δ7Q61L·GMPPNP to map the Cdc42-binding surface.	RESULTS
+126	135	unlabeled	protein_state	A series of 15N HSQC experiments was recorded on 15N-labeled TOCA1 HR1 domain in the presence of increasing concentrations of unlabeled Cdc42Δ7Q61L·GMPPNP to map the Cdc42-binding surface.	RESULTS
+136	154	Cdc42Δ7Q61L·GMPPNP	complex_assembly	A series of 15N HSQC experiments was recorded on 15N-labeled TOCA1 HR1 domain in the presence of increasing concentrations of unlabeled Cdc42Δ7Q61L·GMPPNP to map the Cdc42-binding surface.	RESULTS
+166	187	Cdc42-binding surface	site	A series of 15N HSQC experiments was recorded on 15N-labeled TOCA1 HR1 domain in the presence of increasing concentrations of unlabeled Cdc42Δ7Q61L·GMPPNP to map the Cdc42-binding surface.	RESULTS
+20	28	15N HSQC	experimental_method	A comparison of the 15N HSQC spectra of free HR1 and HR1 in the presence of excess Cdc42 shows that although some peaks were shifted, several were much broader in the complex, and a considerable subset had disappeared (Fig. 4A).	RESULTS
+29	36	spectra	evidence	A comparison of the 15N HSQC spectra of free HR1 and HR1 in the presence of excess Cdc42 shows that although some peaks were shifted, several were much broader in the complex, and a considerable subset had disappeared (Fig. 4A).	RESULTS
+40	44	free	protein_state	A comparison of the 15N HSQC spectra of free HR1 and HR1 in the presence of excess Cdc42 shows that although some peaks were shifted, several were much broader in the complex, and a considerable subset had disappeared (Fig. 4A).	RESULTS
+45	48	HR1	structure_element	A comparison of the 15N HSQC spectra of free HR1 and HR1 in the presence of excess Cdc42 shows that although some peaks were shifted, several were much broader in the complex, and a considerable subset had disappeared (Fig. 4A).	RESULTS
+53	56	HR1	structure_element	A comparison of the 15N HSQC spectra of free HR1 and HR1 in the presence of excess Cdc42 shows that although some peaks were shifted, several were much broader in the complex, and a considerable subset had disappeared (Fig. 4A).	RESULTS
+64	75	presence of	protein_state	A comparison of the 15N HSQC spectra of free HR1 and HR1 in the presence of excess Cdc42 shows that although some peaks were shifted, several were much broader in the complex, and a considerable subset had disappeared (Fig. 4A).	RESULTS
+83	88	Cdc42	protein	A comparison of the 15N HSQC spectra of free HR1 and HR1 in the presence of excess Cdc42 shows that although some peaks were shifted, several were much broader in the complex, and a considerable subset had disappeared (Fig. 4A).	RESULTS
+93	98	Cdc42	protein	This behavior cannot be explained by the increase in molecular mass (from 12 to 33 kDa) when Cdc42 binds and is more likely to be due to conformational exchange.	RESULTS
+8	36	chemical shift perturbations	experimental_method	Overall chemical shift perturbations (CSPs) were calculated for each residue, whereas those that had disappeared were assigned a shift change of 0.2 (Fig. 4B).	RESULTS
+38	42	CSPs	experimental_method	Overall chemical shift perturbations (CSPs) were calculated for each residue, whereas those that had disappeared were assigned a shift change of 0.2 (Fig. 4B).	RESULTS
+33	36	CSP	experimental_method	A peak that disappeared or had a CSP above the mean CSP for the spectrum was considered to be significantly affected.	RESULTS
+52	55	CSP	experimental_method	A peak that disappeared or had a CSP above the mean CSP for the spectrum was considered to be significantly affected.	RESULTS
+12	27	binding surface	site	Mapping the binding surface of Cdc42 onto the TOCA1 HR1 domain.	FIG
+31	36	Cdc42	protein	Mapping the binding surface of Cdc42 onto the TOCA1 HR1 domain.	FIG
+46	51	TOCA1	protein	Mapping the binding surface of Cdc42 onto the TOCA1 HR1 domain.	FIG
+52	55	HR1	structure_element	Mapping the binding surface of Cdc42 onto the TOCA1 HR1 domain.	FIG
+8	16	15N HSQC	experimental_method	 A, the 15N HSQC of 200 μm TOCA1 HR1 domain is shown in the free form (black) and in the presence of a 4-fold molar excess of Cdc42Δ7Q61L·GMPPNP (red).	FIG
+27	32	TOCA1	protein	 A, the 15N HSQC of 200 μm TOCA1 HR1 domain is shown in the free form (black) and in the presence of a 4-fold molar excess of Cdc42Δ7Q61L·GMPPNP (red).	FIG
+33	36	HR1	structure_element	 A, the 15N HSQC of 200 μm TOCA1 HR1 domain is shown in the free form (black) and in the presence of a 4-fold molar excess of Cdc42Δ7Q61L·GMPPNP (red).	FIG
+60	69	free form	protein_state	 A, the 15N HSQC of 200 μm TOCA1 HR1 domain is shown in the free form (black) and in the presence of a 4-fold molar excess of Cdc42Δ7Q61L·GMPPNP (red).	FIG
+89	100	presence of	protein_state	 A, the 15N HSQC of 200 μm TOCA1 HR1 domain is shown in the free form (black) and in the presence of a 4-fold molar excess of Cdc42Δ7Q61L·GMPPNP (red).	FIG
+126	144	Cdc42Δ7Q61L·GMPPNP	complex_assembly	 A, the 15N HSQC of 200 μm TOCA1 HR1 domain is shown in the free form (black) and in the presence of a 4-fold molar excess of Cdc42Δ7Q61L·GMPPNP (red).	FIG
+3	7	CSPs	experimental_method	B, CSPs were calculated as described under “Experimental Procedures” and are shown for backbone and side chain NH groups.	FIG
+9	12	CSP	experimental_method	The mean CSP is marked with a red line.	FIG
+33	44	presence of	protein_state	Residues that disappeared in the presence of Cdc42 were assigned a CSP of 0.2 but were excluded when calculating the mean CSP and are indicated with open bars.	FIG
+45	50	Cdc42	protein	Residues that disappeared in the presence of Cdc42 were assigned a CSP of 0.2 but were excluded when calculating the mean CSP and are indicated with open bars.	FIG
+67	70	CSP	experimental_method	Residues that disappeared in the presence of Cdc42 were assigned a CSP of 0.2 but were excluded when calculating the mean CSP and are indicated with open bars.	FIG
+122	125	CSP	experimental_method	Residues that disappeared in the presence of Cdc42 were assigned a CSP of 0.2 but were excluded when calculating the mean CSP and are indicated with open bars.	FIG
+70	73	CSP	experimental_method	Those that were not traceable due to spectral overlap were assigned a CSP of zero and are marked with an asterisk below the bar.	FIG
+34	38	CSPs	experimental_method	Residues with affected side chain CSPs derived from 13C HSQCs are marked with green asterisks above the bars.	FIG
+52	61	13C HSQCs	experimental_method	Residues with affected side chain CSPs derived from 13C HSQCs are marked with green asterisks above the bars.	FIG
+37	40	HR1	structure_element	C, a schematic representation of the HR1 domain.	FIG
+81	92	Cdc42 bound	protein_state	Residues with significantly affected backbone or side chain chemical shifts when Cdc42 bound and that are buried are colored dark blue, whereas those that are solvent-accessible are colored yellow.	FIG
+159	177	solvent-accessible	protein_state	Residues with significantly affected backbone or side chain chemical shifts when Cdc42 bound and that are buried are colored dark blue, whereas those that are solvent-accessible are colored yellow.	FIG
+77	95	solvent-accessible	protein_state	Residues with significantly affected backbone and side chain groups that are solvent-accessible are colored red.	FIG
+18	32	binding region	site	A close-up of the binding region is shown, with affected side chain heavy atoms shown as sticks.	FIG
+7	31	G protein-binding region	site	D, the G protein-binding region is marked in red onto structures of the HR1 domains as indicated.	FIG
+54	64	structures	evidence	D, the G protein-binding region is marked in red onto structures of the HR1 domains as indicated.	FIG
+72	75	HR1	structure_element	D, the G protein-binding region is marked in red onto structures of the HR1 domains as indicated.	FIG
+0	22	15N HSQC shift mapping	experimental_method	15N HSQC shift mapping experiments report on changes to amide groups, which are mainly inaccessible because they are buried inside the helices and are involved in hydrogen bonds.	RESULTS
+135	142	helices	structure_element	15N HSQC shift mapping experiments report on changes to amide groups, which are mainly inaccessible because they are buried inside the helices and are involved in hydrogen bonds.	RESULTS
+11	19	13C HSQC	experimental_method	Therefore, 13C HSQC and methyl-selective SOFAST-HMQC experiments were also recorded on 15N,13C-labeled TOCA1 HR1 to yield more information on side chain involvement.	RESULTS
+24	52	methyl-selective SOFAST-HMQC	experimental_method	Therefore, 13C HSQC and methyl-selective SOFAST-HMQC experiments were also recorded on 15N,13C-labeled TOCA1 HR1 to yield more information on side chain involvement.	RESULTS
+87	90	15N	chemical	Therefore, 13C HSQC and methyl-selective SOFAST-HMQC experiments were also recorded on 15N,13C-labeled TOCA1 HR1 to yield more information on side chain involvement.	RESULTS
+91	94	13C	chemical	Therefore, 13C HSQC and methyl-selective SOFAST-HMQC experiments were also recorded on 15N,13C-labeled TOCA1 HR1 to yield more information on side chain involvement.	RESULTS
+95	102	labeled	protein_state	Therefore, 13C HSQC and methyl-selective SOFAST-HMQC experiments were also recorded on 15N,13C-labeled TOCA1 HR1 to yield more information on side chain involvement.	RESULTS
+103	108	TOCA1	protein	Therefore, 13C HSQC and methyl-selective SOFAST-HMQC experiments were also recorded on 15N,13C-labeled TOCA1 HR1 to yield more information on side chain involvement.	RESULTS
+109	112	HR1	structure_element	Therefore, 13C HSQC and methyl-selective SOFAST-HMQC experiments were also recorded on 15N,13C-labeled TOCA1 HR1 to yield more information on side chain involvement.	RESULTS
+47	58	presence of	protein_state	Side chains whose CH groups disappeared in the presence of Cdc42 are marked on the graph in Fig. 4B with green asterisks.	RESULTS
+59	64	Cdc42	protein	Side chains whose CH groups disappeared in the presence of Cdc42 are marked on the graph in Fig. 4B with green asterisks.	RESULTS
+0	5	TOCA1	protein	TOCA1 residues whose signals were affected by Cdc42 binding were mapped onto the structure of TOCA1 HR1 (Fig. 4C).	RESULTS
+46	51	Cdc42	protein	TOCA1 residues whose signals were affected by Cdc42 binding were mapped onto the structure of TOCA1 HR1 (Fig. 4C).	RESULTS
+81	90	structure	evidence	TOCA1 residues whose signals were affected by Cdc42 binding were mapped onto the structure of TOCA1 HR1 (Fig. 4C).	RESULTS
+94	99	TOCA1	protein	TOCA1 residues whose signals were affected by Cdc42 binding were mapped onto the structure of TOCA1 HR1 (Fig. 4C).	RESULTS
+100	103	HR1	structure_element	TOCA1 residues whose signals were affected by Cdc42 binding were mapped onto the structure of TOCA1 HR1 (Fig. 4C).	RESULTS
+45	56	coiled-coil	structure_element	The changes were localized to one end of the coiled-coil, and the binding site appeared to include residues from both α-helices and the loop region that joins them.	RESULTS
+66	78	binding site	site	The changes were localized to one end of the coiled-coil, and the binding site appeared to include residues from both α-helices and the loop region that joins them.	RESULTS
+118	127	α-helices	structure_element	The changes were localized to one end of the coiled-coil, and the binding site appeared to include residues from both α-helices and the loop region that joins them.	RESULTS
+136	147	loop region	structure_element	The changes were localized to one end of the coiled-coil, and the binding site appeared to include residues from both α-helices and the loop region that joins them.	RESULTS
+20	37	interhelical loop	structure_element	The residues in the interhelical loop and helix 1 that contact each other (Fig. 3D) show shift changes in their backbone NH and side chains in the presence of Cdc42.	RESULTS
+42	49	helix 1	structure_element	The residues in the interhelical loop and helix 1 that contact each other (Fig. 3D) show shift changes in their backbone NH and side chains in the presence of Cdc42.	RESULTS
+147	158	presence of	protein_state	The residues in the interhelical loop and helix 1 that contact each other (Fig. 3D) show shift changes in their backbone NH and side chains in the presence of Cdc42.	RESULTS
+159	164	Cdc42	protein	The residues in the interhelical loop and helix 1 that contact each other (Fig. 3D) show shift changes in their backbone NH and side chains in the presence of Cdc42.	RESULTS
+31	38	Asn-380	residue_name_number	For example, the side chain of Asn-380 and the backbones of Val-376 and Tyr-377 were significantly affected but are all buried in the free TOCA1 HR1 structure, indicating that local conformational changes in the loop may facilitate complex formation.	RESULTS
+60	67	Val-376	residue_name_number	For example, the side chain of Asn-380 and the backbones of Val-376 and Tyr-377 were significantly affected but are all buried in the free TOCA1 HR1 structure, indicating that local conformational changes in the loop may facilitate complex formation.	RESULTS
+72	79	Tyr-377	residue_name_number	For example, the side chain of Asn-380 and the backbones of Val-376 and Tyr-377 were significantly affected but are all buried in the free TOCA1 HR1 structure, indicating that local conformational changes in the loop may facilitate complex formation.	RESULTS
+134	138	free	protein_state	For example, the side chain of Asn-380 and the backbones of Val-376 and Tyr-377 were significantly affected but are all buried in the free TOCA1 HR1 structure, indicating that local conformational changes in the loop may facilitate complex formation.	RESULTS
+139	144	TOCA1	protein	For example, the side chain of Asn-380 and the backbones of Val-376 and Tyr-377 were significantly affected but are all buried in the free TOCA1 HR1 structure, indicating that local conformational changes in the loop may facilitate complex formation.	RESULTS
+145	148	HR1	structure_element	For example, the side chain of Asn-380 and the backbones of Val-376 and Tyr-377 were significantly affected but are all buried in the free TOCA1 HR1 structure, indicating that local conformational changes in the loop may facilitate complex formation.	RESULTS
+149	158	structure	evidence	For example, the side chain of Asn-380 and the backbones of Val-376 and Tyr-377 were significantly affected but are all buried in the free TOCA1 HR1 structure, indicating that local conformational changes in the loop may facilitate complex formation.	RESULTS
+212	216	loop	structure_element	For example, the side chain of Asn-380 and the backbones of Val-376 and Tyr-377 were significantly affected but are all buried in the free TOCA1 HR1 structure, indicating that local conformational changes in the loop may facilitate complex formation.	RESULTS
+4	26	chemical shift mapping	experimental_method	The chemical shift mapping data indicate that the G protein-binding region of the TOCA1 HR1 domain is broadly similar to that of the CIP4 and PRK1 HR1 domains (Figs. 3B and 4D).	RESULTS
+50	74	G protein-binding region	site	The chemical shift mapping data indicate that the G protein-binding region of the TOCA1 HR1 domain is broadly similar to that of the CIP4 and PRK1 HR1 domains (Figs. 3B and 4D).	RESULTS
+82	87	TOCA1	protein	The chemical shift mapping data indicate that the G protein-binding region of the TOCA1 HR1 domain is broadly similar to that of the CIP4 and PRK1 HR1 domains (Figs. 3B and 4D).	RESULTS
+88	91	HR1	structure_element	The chemical shift mapping data indicate that the G protein-binding region of the TOCA1 HR1 domain is broadly similar to that of the CIP4 and PRK1 HR1 domains (Figs. 3B and 4D).	RESULTS
+133	137	CIP4	protein	The chemical shift mapping data indicate that the G protein-binding region of the TOCA1 HR1 domain is broadly similar to that of the CIP4 and PRK1 HR1 domains (Figs. 3B and 4D).	RESULTS
+142	146	PRK1	protein	The chemical shift mapping data indicate that the G protein-binding region of the TOCA1 HR1 domain is broadly similar to that of the CIP4 and PRK1 HR1 domains (Figs. 3B and 4D).	RESULTS
+147	150	HR1	structure_element	The chemical shift mapping data indicate that the G protein-binding region of the TOCA1 HR1 domain is broadly similar to that of the CIP4 and PRK1 HR1 domains (Figs. 3B and 4D).	RESULTS
+18	21	15N	experimental_method	The corresponding 15N and 13C NMR experiments were also recorded on 15N-Cdc42Δ7Q61L·GMPPNP or 15N/13C -Cdc42Δ7Q61L·GMPPNP in the presence of unlabeled HR1 domain.	RESULTS
+26	33	13C NMR	experimental_method	The corresponding 15N and 13C NMR experiments were also recorded on 15N-Cdc42Δ7Q61L·GMPPNP or 15N/13C -Cdc42Δ7Q61L·GMPPNP in the presence of unlabeled HR1 domain.	RESULTS
+68	71	15N	chemical	The corresponding 15N and 13C NMR experiments were also recorded on 15N-Cdc42Δ7Q61L·GMPPNP or 15N/13C -Cdc42Δ7Q61L·GMPPNP in the presence of unlabeled HR1 domain.	RESULTS
+72	90	Cdc42Δ7Q61L·GMPPNP	complex_assembly	The corresponding 15N and 13C NMR experiments were also recorded on 15N-Cdc42Δ7Q61L·GMPPNP or 15N/13C -Cdc42Δ7Q61L·GMPPNP in the presence of unlabeled HR1 domain.	RESULTS
+94	97	15N	chemical	The corresponding 15N and 13C NMR experiments were also recorded on 15N-Cdc42Δ7Q61L·GMPPNP or 15N/13C -Cdc42Δ7Q61L·GMPPNP in the presence of unlabeled HR1 domain.	RESULTS
+98	101	13C	chemical	The corresponding 15N and 13C NMR experiments were also recorded on 15N-Cdc42Δ7Q61L·GMPPNP or 15N/13C -Cdc42Δ7Q61L·GMPPNP in the presence of unlabeled HR1 domain.	RESULTS
+103	121	Cdc42Δ7Q61L·GMPPNP	complex_assembly	The corresponding 15N and 13C NMR experiments were also recorded on 15N-Cdc42Δ7Q61L·GMPPNP or 15N/13C -Cdc42Δ7Q61L·GMPPNP in the presence of unlabeled HR1 domain.	RESULTS
+129	140	presence of	protein_state	The corresponding 15N and 13C NMR experiments were also recorded on 15N-Cdc42Δ7Q61L·GMPPNP or 15N/13C -Cdc42Δ7Q61L·GMPPNP in the presence of unlabeled HR1 domain.	RESULTS
+141	150	unlabeled	protein_state	The corresponding 15N and 13C NMR experiments were also recorded on 15N-Cdc42Δ7Q61L·GMPPNP or 15N/13C -Cdc42Δ7Q61L·GMPPNP in the presence of unlabeled HR1 domain.	RESULTS
+151	154	HR1	structure_element	The corresponding 15N and 13C NMR experiments were also recorded on 15N-Cdc42Δ7Q61L·GMPPNP or 15N/13C -Cdc42Δ7Q61L·GMPPNP in the presence of unlabeled HR1 domain.	RESULTS
+12	15	CSP	experimental_method	The overall CSP was calculated for each residue.	RESULTS
+21	28	labeled	protein_state	As was the case when labeled HR1 was observed, several peaks were shifted in the complex, but many disappeared, indicating exchange on an unfavorable, millisecond time scale (Fig. 5A).	RESULTS
+29	32	HR1	structure_element	As was the case when labeled HR1 was observed, several peaks were shifted in the complex, but many disappeared, indicating exchange on an unfavorable, millisecond time scale (Fig. 5A).	RESULTS
+93	115	constant time 13C HSQC	experimental_method	Detailed side chain data could not be obtained for all residues due to spectral overlap, but constant time 13C HSQC and methyl-selective SOFAST-HMQC experiments provided further information on certain well resolved side chains (marked with green asterisks in Fig. 5B).	RESULTS
+120	148	methyl-selective SOFAST-HMQC	experimental_method	Detailed side chain data could not be obtained for all residues due to spectral overlap, but constant time 13C HSQC and methyl-selective SOFAST-HMQC experiments provided further information on certain well resolved side chains (marked with green asterisks in Fig. 5B).	RESULTS
+12	27	binding surface	site	Mapping the binding surface of the HR1 domain onto Cdc42.	FIG
+35	38	HR1	structure_element	Mapping the binding surface of the HR1 domain onto Cdc42.	FIG
+51	56	Cdc42	protein	Mapping the binding surface of the HR1 domain onto Cdc42.	FIG
+8	16	15N HSQC	experimental_method	 A, the 15N HSQC of Cdc42Δ7Q61L·GMPPNP is shown in its free form (black) and in the presence of excess TOCA1 HR1 domain (1:2.2, red).	FIG
+20	38	Cdc42Δ7Q61L·GMPPNP	complex_assembly	 A, the 15N HSQC of Cdc42Δ7Q61L·GMPPNP is shown in its free form (black) and in the presence of excess TOCA1 HR1 domain (1:2.2, red).	FIG
+55	64	free form	protein_state	 A, the 15N HSQC of Cdc42Δ7Q61L·GMPPNP is shown in its free form (black) and in the presence of excess TOCA1 HR1 domain (1:2.2, red).	FIG
+84	95	presence of	protein_state	 A, the 15N HSQC of Cdc42Δ7Q61L·GMPPNP is shown in its free form (black) and in the presence of excess TOCA1 HR1 domain (1:2.2, red).	FIG
+103	108	TOCA1	protein	 A, the 15N HSQC of Cdc42Δ7Q61L·GMPPNP is shown in its free form (black) and in the presence of excess TOCA1 HR1 domain (1:2.2, red).	FIG
+109	112	HR1	structure_element	 A, the 15N HSQC of Cdc42Δ7Q61L·GMPPNP is shown in its free form (black) and in the presence of excess TOCA1 HR1 domain (1:2.2, red).	FIG
+3	7	CSPs	experimental_method	B, CSPs are shown for backbone NH groups.	FIG
+32	35	CSP	experimental_method	The red line indicates the mean CSP, plus one S.D. Residues that disappeared in the presence of Cdc42 were assigned a CSP of 0.1 and are indicated with open bars.	FIG
+84	95	presence of	protein_state	The red line indicates the mean CSP, plus one S.D. Residues that disappeared in the presence of Cdc42 were assigned a CSP of 0.1 and are indicated with open bars.	FIG
+96	101	Cdc42	protein	The red line indicates the mean CSP, plus one S.D. Residues that disappeared in the presence of Cdc42 were assigned a CSP of 0.1 and are indicated with open bars.	FIG
+118	121	CSP	experimental_method	The red line indicates the mean CSP, plus one S.D. Residues that disappeared in the presence of Cdc42 were assigned a CSP of 0.1 and are indicated with open bars.	FIG
+35	43	13C HSQC	experimental_method	Residues with disappeared peaks in 13C HSQC experiments are marked on the chart with green asterisks.	FIG
+97	100	NMR	experimental_method	C, the residues with significantly affected backbone and side chain groups are highlighted on an NMR structure of free Cdc42Δ7Q61L·GMPPNP; those that are buried are colored dark blue, whereas those that are solvent-accessible are colored red.	FIG
+101	110	structure	evidence	C, the residues with significantly affected backbone and side chain groups are highlighted on an NMR structure of free Cdc42Δ7Q61L·GMPPNP; those that are buried are colored dark blue, whereas those that are solvent-accessible are colored red.	FIG
+114	118	free	protein_state	C, the residues with significantly affected backbone and side chain groups are highlighted on an NMR structure of free Cdc42Δ7Q61L·GMPPNP; those that are buried are colored dark blue, whereas those that are solvent-accessible are colored red.	FIG
+119	137	Cdc42Δ7Q61L·GMPPNP	complex_assembly	C, the residues with significantly affected backbone and side chain groups are highlighted on an NMR structure of free Cdc42Δ7Q61L·GMPPNP; those that are buried are colored dark blue, whereas those that are solvent-accessible are colored red.	FIG
+207	225	solvent-accessible	protein_state	C, the residues with significantly affected backbone and side chain groups are highlighted on an NMR structure of free Cdc42Δ7Q61L·GMPPNP; those that are buried are colored dark blue, whereas those that are solvent-accessible are colored red.	FIG
+101	119	solvent-accessible	protein_state	Residues with either side chain or backbone groups affected are colored blue if buried and yellow if solvent-accessible.	FIG
+34	47	shift mapping	experimental_method	Residues without information from shift mapping are colored gray.	FIG
+4	12	flexible	protein_state	The flexible switch regions are circled.	FIG
+13	27	switch regions	site	The flexible switch regions are circled.	FIG
+38	64	mean chemical shift change	evidence	As many of the peaks disappeared, the mean chemical shift change was relatively low, so a threshold of the mean plus one S.D. value was used to define a significant CSP.	RESULTS
+165	168	CSP	experimental_method	As many of the peaks disappeared, the mean chemical shift change was relatively low, so a threshold of the mean plus one S.D. value was used to define a significant CSP.	RESULTS
+13	27	switch regions	site	Parts of the switch regions (Fig. 5, B and C) are invisible in NMR spectra recorded on free Cdc42 due to conformational exchange.	RESULTS
+63	66	NMR	experimental_method	Parts of the switch regions (Fig. 5, B and C) are invisible in NMR spectra recorded on free Cdc42 due to conformational exchange.	RESULTS
+67	74	spectra	evidence	Parts of the switch regions (Fig. 5, B and C) are invisible in NMR spectra recorded on free Cdc42 due to conformational exchange.	RESULTS
+87	91	free	protein_state	Parts of the switch regions (Fig. 5, B and C) are invisible in NMR spectra recorded on free Cdc42 due to conformational exchange.	RESULTS
+92	97	Cdc42	protein	Parts of the switch regions (Fig. 5, B and C) are invisible in NMR spectra recorded on free Cdc42 due to conformational exchange.	RESULTS
+6	20	switch regions	site	These switch regions become visible in Cdc42 and other small G protein·effector complexes due to a decrease in conformational freedom upon complex formation.	RESULTS
+39	44	Cdc42	protein	These switch regions become visible in Cdc42 and other small G protein·effector complexes due to a decrease in conformational freedom upon complex formation.	RESULTS
+61	70	G protein	protein_type	These switch regions become visible in Cdc42 and other small G protein·effector complexes due to a decrease in conformational freedom upon complex formation.	RESULTS
+4	18	switch regions	site	The switch regions of Cdc42 did not, however, become visible in the presence of the TOCA1 HR1 domain.	RESULTS
+22	27	Cdc42	protein	The switch regions of Cdc42 did not, however, become visible in the presence of the TOCA1 HR1 domain.	RESULTS
+68	79	presence of	protein_state	The switch regions of Cdc42 did not, however, become visible in the presence of the TOCA1 HR1 domain.	RESULTS
+84	89	TOCA1	protein	The switch regions of Cdc42 did not, however, become visible in the presence of the TOCA1 HR1 domain.	RESULTS
+90	93	HR1	structure_element	The switch regions of Cdc42 did not, however, become visible in the presence of the TOCA1 HR1 domain.	RESULTS
+8	14	Ser-30	residue_name_number	Indeed, Ser-30 of switch I and Arg-66, Arg-68, Leu-70, and Ser-71 of switch II are visible in free Cdc42 but disappear in the presence of the HR1 domain.	RESULTS
+18	26	switch I	site	Indeed, Ser-30 of switch I and Arg-66, Arg-68, Leu-70, and Ser-71 of switch II are visible in free Cdc42 but disappear in the presence of the HR1 domain.	RESULTS
+31	37	Arg-66	residue_name_number	Indeed, Ser-30 of switch I and Arg-66, Arg-68, Leu-70, and Ser-71 of switch II are visible in free Cdc42 but disappear in the presence of the HR1 domain.	RESULTS
+39	45	Arg-68	residue_name_number	Indeed, Ser-30 of switch I and Arg-66, Arg-68, Leu-70, and Ser-71 of switch II are visible in free Cdc42 but disappear in the presence of the HR1 domain.	RESULTS
+47	53	Leu-70	residue_name_number	Indeed, Ser-30 of switch I and Arg-66, Arg-68, Leu-70, and Ser-71 of switch II are visible in free Cdc42 but disappear in the presence of the HR1 domain.	RESULTS
+59	65	Ser-71	residue_name_number	Indeed, Ser-30 of switch I and Arg-66, Arg-68, Leu-70, and Ser-71 of switch II are visible in free Cdc42 but disappear in the presence of the HR1 domain.	RESULTS
+69	78	switch II	site	Indeed, Ser-30 of switch I and Arg-66, Arg-68, Leu-70, and Ser-71 of switch II are visible in free Cdc42 but disappear in the presence of the HR1 domain.	RESULTS
+94	98	free	protein_state	Indeed, Ser-30 of switch I and Arg-66, Arg-68, Leu-70, and Ser-71 of switch II are visible in free Cdc42 but disappear in the presence of the HR1 domain.	RESULTS
+99	104	Cdc42	protein	Indeed, Ser-30 of switch I and Arg-66, Arg-68, Leu-70, and Ser-71 of switch II are visible in free Cdc42 but disappear in the presence of the HR1 domain.	RESULTS
+126	137	presence of	protein_state	Indeed, Ser-30 of switch I and Arg-66, Arg-68, Leu-70, and Ser-71 of switch II are visible in free Cdc42 but disappear in the presence of the HR1 domain.	RESULTS
+142	145	HR1	structure_element	Indeed, Ser-30 of switch I and Arg-66, Arg-68, Leu-70, and Ser-71 of switch II are visible in free Cdc42 but disappear in the presence of the HR1 domain.	RESULTS
+23	37	switch regions	site	This suggests that the switch regions are not rigidified in the HR1 complex and are still in conformational exchange.	RESULTS
+64	67	HR1	structure_element	This suggests that the switch regions are not rigidified in the HR1 complex and are still in conformational exchange.	RESULTS
+56	59	NMR	experimental_method	Nevertheless, mapping of the affected residues onto the NMR structure of free Cdc42Δ7Q61L·GMPPNP (Fig. 5C)8 shows that, although they are relatively widespread compared with changes in the HR1 domain, in general, they are on the face of the protein that includes the switches.	RESULTS
+60	69	structure	evidence	Nevertheless, mapping of the affected residues onto the NMR structure of free Cdc42Δ7Q61L·GMPPNP (Fig. 5C)8 shows that, although they are relatively widespread compared with changes in the HR1 domain, in general, they are on the face of the protein that includes the switches.	RESULTS
+73	77	free	protein_state	Nevertheless, mapping of the affected residues onto the NMR structure of free Cdc42Δ7Q61L·GMPPNP (Fig. 5C)8 shows that, although they are relatively widespread compared with changes in the HR1 domain, in general, they are on the face of the protein that includes the switches.	RESULTS
+78	96	Cdc42Δ7Q61L·GMPPNP	complex_assembly	Nevertheless, mapping of the affected residues onto the NMR structure of free Cdc42Δ7Q61L·GMPPNP (Fig. 5C)8 shows that, although they are relatively widespread compared with changes in the HR1 domain, in general, they are on the face of the protein that includes the switches.	RESULTS
+189	192	HR1	structure_element	Nevertheless, mapping of the affected residues onto the NMR structure of free Cdc42Δ7Q61L·GMPPNP (Fig. 5C)8 shows that, although they are relatively widespread compared with changes in the HR1 domain, in general, they are on the face of the protein that includes the switches.	RESULTS
+267	275	switches	site	Nevertheless, mapping of the affected residues onto the NMR structure of free Cdc42Δ7Q61L·GMPPNP (Fig. 5C)8 shows that, although they are relatively widespread compared with changes in the HR1 domain, in general, they are on the face of the protein that includes the switches.	RESULTS
+13	30	binding interface	site	Although the binding interface may be overestimated, this suggests that the switch regions are involved in binding to TOCA1.	RESULTS
+76	90	switch regions	site	Although the binding interface may be overestimated, this suggests that the switch regions are involved in binding to TOCA1.	RESULTS
+118	123	TOCA1	protein	Although the binding interface may be overestimated, this suggests that the switch regions are involved in binding to TOCA1.	RESULTS
+13	28	Cdc42·TOCA1 HR1	complex_assembly	Modeling the Cdc42·TOCA1 HR1 Complex	RESULTS
+4	18	Cdc42·HR1TOCA1	complex_assembly	The Cdc42·HR1TOCA1 complex was not amenable to full structural analysis due to the weak interaction and the extensive exchange broadening seen in the NMR experiments.	RESULTS
+150	153	NMR	experimental_method	The Cdc42·HR1TOCA1 complex was not amenable to full structural analysis due to the weak interaction and the extensive exchange broadening seen in the NMR experiments.	RESULTS
+0	7	HADDOCK	experimental_method	HADDOCK was therefore used to perform rigid body docking based on the structures of free HR1 domain and Cdc42 and ambiguous interaction restraints derived from the titration experiments described above.	RESULTS
+44	56	body docking	experimental_method	HADDOCK was therefore used to perform rigid body docking based on the structures of free HR1 domain and Cdc42 and ambiguous interaction restraints derived from the titration experiments described above.	RESULTS
+70	80	structures	evidence	HADDOCK was therefore used to perform rigid body docking based on the structures of free HR1 domain and Cdc42 and ambiguous interaction restraints derived from the titration experiments described above.	RESULTS
+84	88	free	protein_state	HADDOCK was therefore used to perform rigid body docking based on the structures of free HR1 domain and Cdc42 and ambiguous interaction restraints derived from the titration experiments described above.	RESULTS
+89	92	HR1	structure_element	HADDOCK was therefore used to perform rigid body docking based on the structures of free HR1 domain and Cdc42 and ambiguous interaction restraints derived from the titration experiments described above.	RESULTS
+104	109	Cdc42	protein	HADDOCK was therefore used to perform rigid body docking based on the structures of free HR1 domain and Cdc42 and ambiguous interaction restraints derived from the titration experiments described above.	RESULTS
+164	185	titration experiments	experimental_method	HADDOCK was therefore used to perform rigid body docking based on the structures of free HR1 domain and Cdc42 and ambiguous interaction restraints derived from the titration experiments described above.	RESULTS
+23	26	HR1	structure_element	The orientation of the HR1 domain with respect to Cdc42 cannot be definitively concluded in the absence of unambiguous distance restraints; hence, HADDOCK produced a set of models in which the HR1 domain contacts the same surface on Cdc42 but is in various orientations with respect to Cdc42.	RESULTS
+50	55	Cdc42	protein	The orientation of the HR1 domain with respect to Cdc42 cannot be definitively concluded in the absence of unambiguous distance restraints; hence, HADDOCK produced a set of models in which the HR1 domain contacts the same surface on Cdc42 but is in various orientations with respect to Cdc42.	RESULTS
+147	154	HADDOCK	experimental_method	The orientation of the HR1 domain with respect to Cdc42 cannot be definitively concluded in the absence of unambiguous distance restraints; hence, HADDOCK produced a set of models in which the HR1 domain contacts the same surface on Cdc42 but is in various orientations with respect to Cdc42.	RESULTS
+193	196	HR1	structure_element	The orientation of the HR1 domain with respect to Cdc42 cannot be definitively concluded in the absence of unambiguous distance restraints; hence, HADDOCK produced a set of models in which the HR1 domain contacts the same surface on Cdc42 but is in various orientations with respect to Cdc42.	RESULTS
+233	238	Cdc42	protein	The orientation of the HR1 domain with respect to Cdc42 cannot be definitively concluded in the absence of unambiguous distance restraints; hence, HADDOCK produced a set of models in which the HR1 domain contacts the same surface on Cdc42 but is in various orientations with respect to Cdc42.	RESULTS
+286	291	Cdc42	protein	The orientation of the HR1 domain with respect to Cdc42 cannot be definitively concluded in the absence of unambiguous distance restraints; hence, HADDOCK produced a set of models in which the HR1 domain contacts the same surface on Cdc42 but is in various orientations with respect to Cdc42.	RESULTS
+28	54	root mean square deviation	evidence	The cluster with the lowest root mean square deviation from the lowest energy structure is assumed to be the best model.	RESULTS
+78	87	structure	evidence	The cluster with the lowest root mean square deviation from the lowest energy structure is assumed to be the best model.	RESULTS
+42	45	HR1	structure_element	By these criteria, in the best model, the HR1 domain is in a similar orientation to the HR1a domain of PRK1 bound to RhoA and the HR1b domain bound to Rac1.	RESULTS
+88	92	HR1a	structure_element	By these criteria, in the best model, the HR1 domain is in a similar orientation to the HR1a domain of PRK1 bound to RhoA and the HR1b domain bound to Rac1.	RESULTS
+103	107	PRK1	protein	By these criteria, in the best model, the HR1 domain is in a similar orientation to the HR1a domain of PRK1 bound to RhoA and the HR1b domain bound to Rac1.	RESULTS
+108	116	bound to	protein_state	By these criteria, in the best model, the HR1 domain is in a similar orientation to the HR1a domain of PRK1 bound to RhoA and the HR1b domain bound to Rac1.	RESULTS
+117	121	RhoA	protein	By these criteria, in the best model, the HR1 domain is in a similar orientation to the HR1a domain of PRK1 bound to RhoA and the HR1b domain bound to Rac1.	RESULTS
+130	134	HR1b	structure_element	By these criteria, in the best model, the HR1 domain is in a similar orientation to the HR1a domain of PRK1 bound to RhoA and the HR1b domain bound to Rac1.	RESULTS
+142	150	bound to	protein_state	By these criteria, in the best model, the HR1 domain is in a similar orientation to the HR1a domain of PRK1 bound to RhoA and the HR1b domain bound to Rac1.	RESULTS
+151	155	Rac1	protein	By these criteria, in the best model, the HR1 domain is in a similar orientation to the HR1a domain of PRK1 bound to RhoA and the HR1b domain bound to Rac1.	RESULTS
+75	84	Rac1-HR1b	complex_assembly	A representative model from this cluster is shown in Fig. 6A alongside the Rac1-HR1b structure (PDB code 2RMK) in Fig. 6B.	RESULTS
+85	94	structure	evidence	A representative model from this cluster is shown in Fig. 6A alongside the Rac1-HR1b structure (PDB code 2RMK) in Fig. 6B.	RESULTS
+9	18	Cdc42·HR1	complex_assembly	Model of Cdc42·HR1 complex.	FIG
+34	43	Cdc42·HR1	complex_assembly	 A, a representative model of the Cdc42·HR1 complex from the cluster closest to the lowest energy model produced using HADDOCK.	FIG
+119	126	HADDOCK	experimental_method	 A, a representative model of the Cdc42·HR1 complex from the cluster closest to the lowest energy model produced using HADDOCK.	FIG
+12	17	Cdc42	protein	Residues of Cdc42 that are affected in the presence of the HR1 domain but are not in close proximity to it are colored in red and labeled.	FIG
+43	54	presence of	protein_state	Residues of Cdc42 that are affected in the presence of the HR1 domain but are not in close proximity to it are colored in red and labeled.	FIG
+59	62	HR1	structure_element	Residues of Cdc42 that are affected in the presence of the HR1 domain but are not in close proximity to it are colored in red and labeled.	FIG
+3	12	structure	evidence	B, structure of Rac1 in complex with the HR1b domain of PRK1 (PDB code 2RMK).	FIG
+16	20	Rac1	protein	B, structure of Rac1 in complex with the HR1b domain of PRK1 (PDB code 2RMK).	FIG
+21	36	in complex with	protein_state	B, structure of Rac1 in complex with the HR1b domain of PRK1 (PDB code 2RMK).	FIG
+41	45	HR1b	structure_element	B, structure of Rac1 in complex with the HR1b domain of PRK1 (PDB code 2RMK).	FIG
+56	60	PRK1	protein	B, structure of Rac1 in complex with the HR1b domain of PRK1 (PDB code 2RMK).	FIG
+3	21	sequence alignment	experimental_method	C, sequence alignment of RhoA, Cdc42 and Rac1.	FIG
+25	29	RhoA	protein	C, sequence alignment of RhoA, Cdc42 and Rac1.	FIG
+31	36	Cdc42	protein	C, sequence alignment of RhoA, Cdc42 and Rac1.	FIG
+41	45	Rac1	protein	C, sequence alignment of RhoA, Cdc42 and Rac1.	FIG
+20	24	RhoA	protein	Contact residues of RhoA and Rac1 to PRK1 HR1a and HR1b, respectively, are colored cyan.	FIG
+29	33	Rac1	protein	Contact residues of RhoA and Rac1 to PRK1 HR1a and HR1b, respectively, are colored cyan.	FIG
+37	41	PRK1	protein	Contact residues of RhoA and Rac1 to PRK1 HR1a and HR1b, respectively, are colored cyan.	FIG
+42	46	HR1a	structure_element	Contact residues of RhoA and Rac1 to PRK1 HR1a and HR1b, respectively, are colored cyan.	FIG
+51	55	HR1b	structure_element	Contact residues of RhoA and Rac1 to PRK1 HR1a and HR1b, respectively, are colored cyan.	FIG
+12	17	Cdc42	protein	Residues of Cdc42 that disappear or show chemical shift changes in the presence of TOCA1 are colored cyan if also identified as contacts in RhoA and Rac1 and yellow if they are not.	FIG
+71	82	presence of	protein_state	Residues of Cdc42 that disappear or show chemical shift changes in the presence of TOCA1 are colored cyan if also identified as contacts in RhoA and Rac1 and yellow if they are not.	FIG
+83	88	TOCA1	protein	Residues of Cdc42 that disappear or show chemical shift changes in the presence of TOCA1 are colored cyan if also identified as contacts in RhoA and Rac1 and yellow if they are not.	FIG
+140	144	RhoA	protein	Residues of Cdc42 that disappear or show chemical shift changes in the presence of TOCA1 are colored cyan if also identified as contacts in RhoA and Rac1 and yellow if they are not.	FIG
+149	153	Rac1	protein	Residues of Cdc42 that disappear or show chemical shift changes in the presence of TOCA1 are colored cyan if also identified as contacts in RhoA and Rac1 and yellow if they are not.	FIG
+23	27	Rac1	protein	Residues equivalent to Rac1 and RhoA contact sites but that are invisible in free Cdc42 are gray.	FIG
+32	36	RhoA	protein	Residues equivalent to Rac1 and RhoA contact sites but that are invisible in free Cdc42 are gray.	FIG
+37	50	contact sites	site	Residues equivalent to Rac1 and RhoA contact sites but that are invisible in free Cdc42 are gray.	FIG
+77	81	free	protein_state	Residues equivalent to Rac1 and RhoA contact sites but that are invisible in free Cdc42 are gray.	FIG
+82	87	Cdc42	protein	Residues equivalent to Rac1 and RhoA contact sites but that are invisible in free Cdc42 are gray.	FIG
+30	39	Cdc42·HR1	complex_assembly	D, regions of interest of the Cdc42·HR1 domain model.	FIG
+23	33	structures	evidence	The four lowest energy structures in the chosen HADDOCK cluster are shown overlaid, with the residues of interest shown as sticks and labeled.	FIG
+48	55	HADDOCK	experimental_method	The four lowest energy structures in the chosen HADDOCK cluster are shown overlaid, with the residues of interest shown as sticks and labeled.	FIG
+28	33	TOCA1	protein	Cdc42 is shown in cyan, and TOCA1 is shown in purple.	FIG
+2	20	sequence alignment	experimental_method	A sequence alignment of RhoA, Cdc42, and Rac1 is shown in Fig. 6C.	RESULTS
+24	28	RhoA	protein	A sequence alignment of RhoA, Cdc42, and Rac1 is shown in Fig. 6C.	RESULTS
+30	35	Cdc42	protein	A sequence alignment of RhoA, Cdc42, and Rac1 is shown in Fig. 6C.	RESULTS
+41	45	Rac1	protein	A sequence alignment of RhoA, Cdc42, and Rac1 is shown in Fig. 6C.	RESULTS
+4	8	RhoA	protein	The RhoA and Rac1 contact residues in the switch regions are invisible in the spectra of Cdc42, but they are generally conserved between all three G proteins.	RESULTS
+13	17	Rac1	protein	The RhoA and Rac1 contact residues in the switch regions are invisible in the spectra of Cdc42, but they are generally conserved between all three G proteins.	RESULTS
+42	56	switch regions	site	The RhoA and Rac1 contact residues in the switch regions are invisible in the spectra of Cdc42, but they are generally conserved between all three G proteins.	RESULTS
+78	85	spectra	evidence	The RhoA and Rac1 contact residues in the switch regions are invisible in the spectra of Cdc42, but they are generally conserved between all three G proteins.	RESULTS
+89	94	Cdc42	protein	The RhoA and Rac1 contact residues in the switch regions are invisible in the spectra of Cdc42, but they are generally conserved between all three G proteins.	RESULTS
+119	128	conserved	protein_state	The RhoA and Rac1 contact residues in the switch regions are invisible in the spectra of Cdc42, but they are generally conserved between all three G proteins.	RESULTS
+147	157	G proteins	protein_type	The RhoA and Rac1 contact residues in the switch regions are invisible in the spectra of Cdc42, but they are generally conserved between all three G proteins.	RESULTS
+8	13	Cdc42	protein	Several Cdc42 residues identified by chemical shift mapping are not in close contact in the Cdc42·TOCA1 model (Fig. 6A).	RESULTS
+37	59	chemical shift mapping	experimental_method	Several Cdc42 residues identified by chemical shift mapping are not in close contact in the Cdc42·TOCA1 model (Fig. 6A).	RESULTS
+92	103	Cdc42·TOCA1	complex_assembly	Several Cdc42 residues identified by chemical shift mapping are not in close contact in the Cdc42·TOCA1 model (Fig. 6A).	RESULTS
+48	54	Thr-24	residue_name_number	Some of these can be rationalized; for example, Thr-24Cdc42, Leu-160Cdc42, and Lys-163Cdc42 all pack behind switch I and are likely to be affected by conformational changes within the switch, while Glu-95Cdc42 and Lys-96Cdc42 are in the helix behind switch II.	RESULTS
+54	59	Cdc42	protein	Some of these can be rationalized; for example, Thr-24Cdc42, Leu-160Cdc42, and Lys-163Cdc42 all pack behind switch I and are likely to be affected by conformational changes within the switch, while Glu-95Cdc42 and Lys-96Cdc42 are in the helix behind switch II.	RESULTS
+61	68	Leu-160	residue_name_number	Some of these can be rationalized; for example, Thr-24Cdc42, Leu-160Cdc42, and Lys-163Cdc42 all pack behind switch I and are likely to be affected by conformational changes within the switch, while Glu-95Cdc42 and Lys-96Cdc42 are in the helix behind switch II.	RESULTS
+68	73	Cdc42	protein	Some of these can be rationalized; for example, Thr-24Cdc42, Leu-160Cdc42, and Lys-163Cdc42 all pack behind switch I and are likely to be affected by conformational changes within the switch, while Glu-95Cdc42 and Lys-96Cdc42 are in the helix behind switch II.	RESULTS
+79	86	Lys-163	residue_name_number	Some of these can be rationalized; for example, Thr-24Cdc42, Leu-160Cdc42, and Lys-163Cdc42 all pack behind switch I and are likely to be affected by conformational changes within the switch, while Glu-95Cdc42 and Lys-96Cdc42 are in the helix behind switch II.	RESULTS
+86	91	Cdc42	protein	Some of these can be rationalized; for example, Thr-24Cdc42, Leu-160Cdc42, and Lys-163Cdc42 all pack behind switch I and are likely to be affected by conformational changes within the switch, while Glu-95Cdc42 and Lys-96Cdc42 are in the helix behind switch II.	RESULTS
+108	116	switch I	site	Some of these can be rationalized; for example, Thr-24Cdc42, Leu-160Cdc42, and Lys-163Cdc42 all pack behind switch I and are likely to be affected by conformational changes within the switch, while Glu-95Cdc42 and Lys-96Cdc42 are in the helix behind switch II.	RESULTS
+184	190	switch	site	Some of these can be rationalized; for example, Thr-24Cdc42, Leu-160Cdc42, and Lys-163Cdc42 all pack behind switch I and are likely to be affected by conformational changes within the switch, while Glu-95Cdc42 and Lys-96Cdc42 are in the helix behind switch II.	RESULTS
+198	204	Glu-95	residue_name_number	Some of these can be rationalized; for example, Thr-24Cdc42, Leu-160Cdc42, and Lys-163Cdc42 all pack behind switch I and are likely to be affected by conformational changes within the switch, while Glu-95Cdc42 and Lys-96Cdc42 are in the helix behind switch II.	RESULTS
+204	209	Cdc42	protein	Some of these can be rationalized; for example, Thr-24Cdc42, Leu-160Cdc42, and Lys-163Cdc42 all pack behind switch I and are likely to be affected by conformational changes within the switch, while Glu-95Cdc42 and Lys-96Cdc42 are in the helix behind switch II.	RESULTS
+214	220	Lys-96	residue_name_number	Some of these can be rationalized; for example, Thr-24Cdc42, Leu-160Cdc42, and Lys-163Cdc42 all pack behind switch I and are likely to be affected by conformational changes within the switch, while Glu-95Cdc42 and Lys-96Cdc42 are in the helix behind switch II.	RESULTS
+220	225	Cdc42	protein	Some of these can be rationalized; for example, Thr-24Cdc42, Leu-160Cdc42, and Lys-163Cdc42 all pack behind switch I and are likely to be affected by conformational changes within the switch, while Glu-95Cdc42 and Lys-96Cdc42 are in the helix behind switch II.	RESULTS
+237	242	helix	structure_element	Some of these can be rationalized; for example, Thr-24Cdc42, Leu-160Cdc42, and Lys-163Cdc42 all pack behind switch I and are likely to be affected by conformational changes within the switch, while Glu-95Cdc42 and Lys-96Cdc42 are in the helix behind switch II.	RESULTS
+250	259	switch II	site	Some of these can be rationalized; for example, Thr-24Cdc42, Leu-160Cdc42, and Lys-163Cdc42 all pack behind switch I and are likely to be affected by conformational changes within the switch, while Glu-95Cdc42 and Lys-96Cdc42 are in the helix behind switch II.	RESULTS
+40	51	Cdc42·TOCA1	complex_assembly	Other residues that are affected in the Cdc42·TOCA1 complex but that do not correspond to contact residues of RhoA or Rac1 (Fig. 6C) include Gln-2Cdc42, Lys-16Cdc42, Thr-52Cdc42, and Arg-68Cdc42.	RESULTS
+110	114	RhoA	protein	Other residues that are affected in the Cdc42·TOCA1 complex but that do not correspond to contact residues of RhoA or Rac1 (Fig. 6C) include Gln-2Cdc42, Lys-16Cdc42, Thr-52Cdc42, and Arg-68Cdc42.	RESULTS
+118	122	Rac1	protein	Other residues that are affected in the Cdc42·TOCA1 complex but that do not correspond to contact residues of RhoA or Rac1 (Fig. 6C) include Gln-2Cdc42, Lys-16Cdc42, Thr-52Cdc42, and Arg-68Cdc42.	RESULTS
+141	146	Gln-2	residue_name_number	Other residues that are affected in the Cdc42·TOCA1 complex but that do not correspond to contact residues of RhoA or Rac1 (Fig. 6C) include Gln-2Cdc42, Lys-16Cdc42, Thr-52Cdc42, and Arg-68Cdc42.	RESULTS
+146	151	Cdc42	protein	Other residues that are affected in the Cdc42·TOCA1 complex but that do not correspond to contact residues of RhoA or Rac1 (Fig. 6C) include Gln-2Cdc42, Lys-16Cdc42, Thr-52Cdc42, and Arg-68Cdc42.	RESULTS
+153	159	Lys-16	residue_name_number	Other residues that are affected in the Cdc42·TOCA1 complex but that do not correspond to contact residues of RhoA or Rac1 (Fig. 6C) include Gln-2Cdc42, Lys-16Cdc42, Thr-52Cdc42, and Arg-68Cdc42.	RESULTS
+159	164	Cdc42	protein	Other residues that are affected in the Cdc42·TOCA1 complex but that do not correspond to contact residues of RhoA or Rac1 (Fig. 6C) include Gln-2Cdc42, Lys-16Cdc42, Thr-52Cdc42, and Arg-68Cdc42.	RESULTS
+166	172	Thr-52	residue_name_number	Other residues that are affected in the Cdc42·TOCA1 complex but that do not correspond to contact residues of RhoA or Rac1 (Fig. 6C) include Gln-2Cdc42, Lys-16Cdc42, Thr-52Cdc42, and Arg-68Cdc42.	RESULTS
+172	177	Cdc42	protein	Other residues that are affected in the Cdc42·TOCA1 complex but that do not correspond to contact residues of RhoA or Rac1 (Fig. 6C) include Gln-2Cdc42, Lys-16Cdc42, Thr-52Cdc42, and Arg-68Cdc42.	RESULTS
+183	189	Arg-68	residue_name_number	Other residues that are affected in the Cdc42·TOCA1 complex but that do not correspond to contact residues of RhoA or Rac1 (Fig. 6C) include Gln-2Cdc42, Lys-16Cdc42, Thr-52Cdc42, and Arg-68Cdc42.	RESULTS
+189	194	Cdc42	protein	Other residues that are affected in the Cdc42·TOCA1 complex but that do not correspond to contact residues of RhoA or Rac1 (Fig. 6C) include Gln-2Cdc42, Lys-16Cdc42, Thr-52Cdc42, and Arg-68Cdc42.	RESULTS
+0	6	Lys-16	residue_name_number	Lys-16Cdc42 is unlikely to be a contact residue because it is involved in nucleotide binding, but the others may represent specific Cdc42-TOCA1 contacts.	RESULTS
+6	11	Cdc42	protein	Lys-16Cdc42 is unlikely to be a contact residue because it is involved in nucleotide binding, but the others may represent specific Cdc42-TOCA1 contacts.	RESULTS
+132	143	Cdc42-TOCA1	complex_assembly	Lys-16Cdc42 is unlikely to be a contact residue because it is involved in nucleotide binding, but the others may represent specific Cdc42-TOCA1 contacts.	RESULTS
+20	26	N-WASP	protein	Competition between N-WASP and TOCA1	RESULTS
+31	36	TOCA1	protein	Competition between N-WASP and TOCA1	RESULTS
+106	111	TOCA1	protein	From the known interactions and effects of the proteins in biological systems, it has been suggested that TOCA1 and N-WASP could bind Cdc42 simultaneously.	RESULTS
+116	122	N-WASP	protein	From the known interactions and effects of the proteins in biological systems, it has been suggested that TOCA1 and N-WASP could bind Cdc42 simultaneously.	RESULTS
+134	139	Cdc42	protein	From the known interactions and effects of the proteins in biological systems, it has been suggested that TOCA1 and N-WASP could bind Cdc42 simultaneously.	RESULTS
+38	56	Cdc42·N-WASP·TOCA1	complex_assembly	Studies in CHO cells indicated that a Cdc42·N-WASP·TOCA1 complex existed because FRET was observed between RFP-TOCA1 and GFP-N-WASP, and the efficiency was decreased when an N-WASP mutant was used that no longer binds Cdc42.	RESULTS
+81	85	FRET	evidence	Studies in CHO cells indicated that a Cdc42·N-WASP·TOCA1 complex existed because FRET was observed between RFP-TOCA1 and GFP-N-WASP, and the efficiency was decreased when an N-WASP mutant was used that no longer binds Cdc42.	RESULTS
+107	110	RFP	chemical	Studies in CHO cells indicated that a Cdc42·N-WASP·TOCA1 complex existed because FRET was observed between RFP-TOCA1 and GFP-N-WASP, and the efficiency was decreased when an N-WASP mutant was used that no longer binds Cdc42.	RESULTS
+111	116	TOCA1	protein	Studies in CHO cells indicated that a Cdc42·N-WASP·TOCA1 complex existed because FRET was observed between RFP-TOCA1 and GFP-N-WASP, and the efficiency was decreased when an N-WASP mutant was used that no longer binds Cdc42.	RESULTS
+121	124	GFP	chemical	Studies in CHO cells indicated that a Cdc42·N-WASP·TOCA1 complex existed because FRET was observed between RFP-TOCA1 and GFP-N-WASP, and the efficiency was decreased when an N-WASP mutant was used that no longer binds Cdc42.	RESULTS
+125	131	N-WASP	protein	Studies in CHO cells indicated that a Cdc42·N-WASP·TOCA1 complex existed because FRET was observed between RFP-TOCA1 and GFP-N-WASP, and the efficiency was decreased when an N-WASP mutant was used that no longer binds Cdc42.	RESULTS
+174	180	N-WASP	protein	Studies in CHO cells indicated that a Cdc42·N-WASP·TOCA1 complex existed because FRET was observed between RFP-TOCA1 and GFP-N-WASP, and the efficiency was decreased when an N-WASP mutant was used that no longer binds Cdc42.	RESULTS
+181	187	mutant	protein_state	Studies in CHO cells indicated that a Cdc42·N-WASP·TOCA1 complex existed because FRET was observed between RFP-TOCA1 and GFP-N-WASP, and the efficiency was decreased when an N-WASP mutant was used that no longer binds Cdc42.	RESULTS
+218	223	Cdc42	protein	Studies in CHO cells indicated that a Cdc42·N-WASP·TOCA1 complex existed because FRET was observed between RFP-TOCA1 and GFP-N-WASP, and the efficiency was decreased when an N-WASP mutant was used that no longer binds Cdc42.	RESULTS
+3	10	overlay	experimental_method	An overlay of the HADDOCK model of the Cdc42·HR1TOCA1 complex and the structure of Cdc42 in complex with the GBD of the N-WASP homologue, WASP (PDB code 1CEE), shows that the HR1 and GBD binding sites only partly overlap, and, therefore, a ternary complex remained possible (Fig. 7A).	RESULTS
+18	25	HADDOCK	experimental_method	An overlay of the HADDOCK model of the Cdc42·HR1TOCA1 complex and the structure of Cdc42 in complex with the GBD of the N-WASP homologue, WASP (PDB code 1CEE), shows that the HR1 and GBD binding sites only partly overlap, and, therefore, a ternary complex remained possible (Fig. 7A).	RESULTS
+26	31	model	evidence	An overlay of the HADDOCK model of the Cdc42·HR1TOCA1 complex and the structure of Cdc42 in complex with the GBD of the N-WASP homologue, WASP (PDB code 1CEE), shows that the HR1 and GBD binding sites only partly overlap, and, therefore, a ternary complex remained possible (Fig. 7A).	RESULTS
+39	53	Cdc42·HR1TOCA1	complex_assembly	An overlay of the HADDOCK model of the Cdc42·HR1TOCA1 complex and the structure of Cdc42 in complex with the GBD of the N-WASP homologue, WASP (PDB code 1CEE), shows that the HR1 and GBD binding sites only partly overlap, and, therefore, a ternary complex remained possible (Fig. 7A).	RESULTS
+70	79	structure	evidence	An overlay of the HADDOCK model of the Cdc42·HR1TOCA1 complex and the structure of Cdc42 in complex with the GBD of the N-WASP homologue, WASP (PDB code 1CEE), shows that the HR1 and GBD binding sites only partly overlap, and, therefore, a ternary complex remained possible (Fig. 7A).	RESULTS
+83	88	Cdc42	protein	An overlay of the HADDOCK model of the Cdc42·HR1TOCA1 complex and the structure of Cdc42 in complex with the GBD of the N-WASP homologue, WASP (PDB code 1CEE), shows that the HR1 and GBD binding sites only partly overlap, and, therefore, a ternary complex remained possible (Fig. 7A).	RESULTS
+89	104	in complex with	protein_state	An overlay of the HADDOCK model of the Cdc42·HR1TOCA1 complex and the structure of Cdc42 in complex with the GBD of the N-WASP homologue, WASP (PDB code 1CEE), shows that the HR1 and GBD binding sites only partly overlap, and, therefore, a ternary complex remained possible (Fig. 7A).	RESULTS
+109	112	GBD	structure_element	An overlay of the HADDOCK model of the Cdc42·HR1TOCA1 complex and the structure of Cdc42 in complex with the GBD of the N-WASP homologue, WASP (PDB code 1CEE), shows that the HR1 and GBD binding sites only partly overlap, and, therefore, a ternary complex remained possible (Fig. 7A).	RESULTS
+120	126	N-WASP	protein	An overlay of the HADDOCK model of the Cdc42·HR1TOCA1 complex and the structure of Cdc42 in complex with the GBD of the N-WASP homologue, WASP (PDB code 1CEE), shows that the HR1 and GBD binding sites only partly overlap, and, therefore, a ternary complex remained possible (Fig. 7A).	RESULTS
+138	142	WASP	protein	An overlay of the HADDOCK model of the Cdc42·HR1TOCA1 complex and the structure of Cdc42 in complex with the GBD of the N-WASP homologue, WASP (PDB code 1CEE), shows that the HR1 and GBD binding sites only partly overlap, and, therefore, a ternary complex remained possible (Fig. 7A).	RESULTS
+175	178	HR1	structure_element	An overlay of the HADDOCK model of the Cdc42·HR1TOCA1 complex and the structure of Cdc42 in complex with the GBD of the N-WASP homologue, WASP (PDB code 1CEE), shows that the HR1 and GBD binding sites only partly overlap, and, therefore, a ternary complex remained possible (Fig. 7A).	RESULTS
+183	200	GBD binding sites	site	An overlay of the HADDOCK model of the Cdc42·HR1TOCA1 complex and the structure of Cdc42 in complex with the GBD of the N-WASP homologue, WASP (PDB code 1CEE), shows that the HR1 and GBD binding sites only partly overlap, and, therefore, a ternary complex remained possible (Fig. 7A).	RESULTS
+19	30	presence of	protein_state	Interestingly, the presence of the TOCA1 HR1 would not prevent the core CRIB of WASP from binding to Cdc42, although the regions C-terminal to the CRIB that are required for high affinity binding of WASP would interfere sterically with the TOCA1 HR1.	RESULTS
+35	40	TOCA1	protein	Interestingly, the presence of the TOCA1 HR1 would not prevent the core CRIB of WASP from binding to Cdc42, although the regions C-terminal to the CRIB that are required for high affinity binding of WASP would interfere sterically with the TOCA1 HR1.	RESULTS
+41	44	HR1	structure_element	Interestingly, the presence of the TOCA1 HR1 would not prevent the core CRIB of WASP from binding to Cdc42, although the regions C-terminal to the CRIB that are required for high affinity binding of WASP would interfere sterically with the TOCA1 HR1.	RESULTS
+72	76	CRIB	structure_element	Interestingly, the presence of the TOCA1 HR1 would not prevent the core CRIB of WASP from binding to Cdc42, although the regions C-terminal to the CRIB that are required for high affinity binding of WASP would interfere sterically with the TOCA1 HR1.	RESULTS
+80	84	WASP	protein	Interestingly, the presence of the TOCA1 HR1 would not prevent the core CRIB of WASP from binding to Cdc42, although the regions C-terminal to the CRIB that are required for high affinity binding of WASP would interfere sterically with the TOCA1 HR1.	RESULTS
+101	106	Cdc42	protein	Interestingly, the presence of the TOCA1 HR1 would not prevent the core CRIB of WASP from binding to Cdc42, although the regions C-terminal to the CRIB that are required for high affinity binding of WASP would interfere sterically with the TOCA1 HR1.	RESULTS
+147	151	CRIB	structure_element	Interestingly, the presence of the TOCA1 HR1 would not prevent the core CRIB of WASP from binding to Cdc42, although the regions C-terminal to the CRIB that are required for high affinity binding of WASP would interfere sterically with the TOCA1 HR1.	RESULTS
+199	203	WASP	protein	Interestingly, the presence of the TOCA1 HR1 would not prevent the core CRIB of WASP from binding to Cdc42, although the regions C-terminal to the CRIB that are required for high affinity binding of WASP would interfere sterically with the TOCA1 HR1.	RESULTS
+240	245	TOCA1	protein	Interestingly, the presence of the TOCA1 HR1 would not prevent the core CRIB of WASP from binding to Cdc42, although the regions C-terminal to the CRIB that are required for high affinity binding of WASP would interfere sterically with the TOCA1 HR1.	RESULTS
+246	249	HR1	structure_element	Interestingly, the presence of the TOCA1 HR1 would not prevent the core CRIB of WASP from binding to Cdc42, although the regions C-terminal to the CRIB that are required for high affinity binding of WASP would interfere sterically with the TOCA1 HR1.	RESULTS
+18	22	WASP	protein	A basic region in WASP including three lysines (residues 230–232), N-terminal to the core CRIB, has been implicated in an electrostatic steering mechanism, and these residues would be free to bind in the presence of TOCA1 HR1 (Fig. 7A).	RESULTS
+39	46	lysines	residue_name	A basic region in WASP including three lysines (residues 230–232), N-terminal to the core CRIB, has been implicated in an electrostatic steering mechanism, and these residues would be free to bind in the presence of TOCA1 HR1 (Fig. 7A).	RESULTS
+57	64	230–232	residue_range	A basic region in WASP including three lysines (residues 230–232), N-terminal to the core CRIB, has been implicated in an electrostatic steering mechanism, and these residues would be free to bind in the presence of TOCA1 HR1 (Fig. 7A).	RESULTS
+90	94	CRIB	structure_element	A basic region in WASP including three lysines (residues 230–232), N-terminal to the core CRIB, has been implicated in an electrostatic steering mechanism, and these residues would be free to bind in the presence of TOCA1 HR1 (Fig. 7A).	RESULTS
+204	215	presence of	protein_state	A basic region in WASP including three lysines (residues 230–232), N-terminal to the core CRIB, has been implicated in an electrostatic steering mechanism, and these residues would be free to bind in the presence of TOCA1 HR1 (Fig. 7A).	RESULTS
+216	221	TOCA1	protein	A basic region in WASP including three lysines (residues 230–232), N-terminal to the core CRIB, has been implicated in an electrostatic steering mechanism, and these residues would be free to bind in the presence of TOCA1 HR1 (Fig. 7A).	RESULTS
+222	225	HR1	structure_element	A basic region in WASP including three lysines (residues 230–232), N-terminal to the core CRIB, has been implicated in an electrostatic steering mechanism, and these residues would be free to bind in the presence of TOCA1 HR1 (Fig. 7A).	RESULTS
+4	10	N-WASP	protein	The N-WASP GBD displaces the TOCA1 HR1 domain.	FIG
+11	14	GBD	structure_element	The N-WASP GBD displaces the TOCA1 HR1 domain.	FIG
+29	34	TOCA1	protein	The N-WASP GBD displaces the TOCA1 HR1 domain.	FIG
+35	38	HR1	structure_element	The N-WASP GBD displaces the TOCA1 HR1 domain.	FIG
+21	32	Cdc42·TOCA1	complex_assembly	 A, the model of the Cdc42·TOCA1 HR1 domain complex overlaid with the Cdc42-WASP structure.	FIG
+33	36	HR1	structure_element	 A, the model of the Cdc42·TOCA1 HR1 domain complex overlaid with the Cdc42-WASP structure.	FIG
+70	80	Cdc42-WASP	complex_assembly	 A, the model of the Cdc42·TOCA1 HR1 domain complex overlaid with the Cdc42-WASP structure.	FIG
+81	90	structure	evidence	 A, the model of the Cdc42·TOCA1 HR1 domain complex overlaid with the Cdc42-WASP structure.	FIG
+29	34	TOCA1	protein	Cdc42 is shown in green, and TOCA1 is shown in purple.	FIG
+9	13	CRIB	structure_element	The core CRIB region of WASP is shown in red, whereas its basic region is shown in orange and the C-terminal region required for maximal affinity is shown in cyan.	FIG
+24	28	WASP	protein	The core CRIB region of WASP is shown in red, whereas its basic region is shown in orange and the C-terminal region required for maximal affinity is shown in cyan.	FIG
+44	49	Cdc42	protein	A semitransparent surface representation of Cdc42 and WASP is shown overlaid with the schematic.	FIG
+54	58	WASP	protein	A semitransparent surface representation of Cdc42 and WASP is shown overlaid with the schematic.	FIG
+3	18	competition SPA	experimental_method	B, competition SPA experiments carried out with indicated concentrations of the N-WASP GBD construct titrated into 30 nm GST-ACK or GST-WASP GBD and 30 nm Cdc42Δ7Q61L·[3H]GTP.	FIG
+80	86	N-WASP	protein	B, competition SPA experiments carried out with indicated concentrations of the N-WASP GBD construct titrated into 30 nm GST-ACK or GST-WASP GBD and 30 nm Cdc42Δ7Q61L·[3H]GTP.	FIG
+87	90	GBD	structure_element	B, competition SPA experiments carried out with indicated concentrations of the N-WASP GBD construct titrated into 30 nm GST-ACK or GST-WASP GBD and 30 nm Cdc42Δ7Q61L·[3H]GTP.	FIG
+101	109	titrated	experimental_method	B, competition SPA experiments carried out with indicated concentrations of the N-WASP GBD construct titrated into 30 nm GST-ACK or GST-WASP GBD and 30 nm Cdc42Δ7Q61L·[3H]GTP.	FIG
+121	128	GST-ACK	mutant	B, competition SPA experiments carried out with indicated concentrations of the N-WASP GBD construct titrated into 30 nm GST-ACK or GST-WASP GBD and 30 nm Cdc42Δ7Q61L·[3H]GTP.	FIG
+132	140	GST-WASP	mutant	B, competition SPA experiments carried out with indicated concentrations of the N-WASP GBD construct titrated into 30 nm GST-ACK or GST-WASP GBD and 30 nm Cdc42Δ7Q61L·[3H]GTP.	FIG
+141	144	GBD	structure_element	B, competition SPA experiments carried out with indicated concentrations of the N-WASP GBD construct titrated into 30 nm GST-ACK or GST-WASP GBD and 30 nm Cdc42Δ7Q61L·[3H]GTP.	FIG
+155	174	Cdc42Δ7Q61L·[3H]GTP	complex_assembly	B, competition SPA experiments carried out with indicated concentrations of the N-WASP GBD construct titrated into 30 nm GST-ACK or GST-WASP GBD and 30 nm Cdc42Δ7Q61L·[3H]GTP.	FIG
+27	35	15N HSQC	experimental_method	C, Selected regions of the 15N HSQC of 145 μm Cdc42Δ7Q61L·GMPPNP with the indicated ratios of the TOCA1 HR1 domain, the N-WASP GBD, or both, showing that the TOCA HR1 domain does not displace the N-WASP GBD.	FIG
+46	64	Cdc42Δ7Q61L·GMPPNP	complex_assembly	C, Selected regions of the 15N HSQC of 145 μm Cdc42Δ7Q61L·GMPPNP with the indicated ratios of the TOCA1 HR1 domain, the N-WASP GBD, or both, showing that the TOCA HR1 domain does not displace the N-WASP GBD.	FIG
+98	103	TOCA1	protein	C, Selected regions of the 15N HSQC of 145 μm Cdc42Δ7Q61L·GMPPNP with the indicated ratios of the TOCA1 HR1 domain, the N-WASP GBD, or both, showing that the TOCA HR1 domain does not displace the N-WASP GBD.	FIG
+104	107	HR1	structure_element	C, Selected regions of the 15N HSQC of 145 μm Cdc42Δ7Q61L·GMPPNP with the indicated ratios of the TOCA1 HR1 domain, the N-WASP GBD, or both, showing that the TOCA HR1 domain does not displace the N-WASP GBD.	FIG
+120	126	N-WASP	protein	C, Selected regions of the 15N HSQC of 145 μm Cdc42Δ7Q61L·GMPPNP with the indicated ratios of the TOCA1 HR1 domain, the N-WASP GBD, or both, showing that the TOCA HR1 domain does not displace the N-WASP GBD.	FIG
+127	130	GBD	structure_element	C, Selected regions of the 15N HSQC of 145 μm Cdc42Δ7Q61L·GMPPNP with the indicated ratios of the TOCA1 HR1 domain, the N-WASP GBD, or both, showing that the TOCA HR1 domain does not displace the N-WASP GBD.	FIG
+158	162	TOCA	protein	C, Selected regions of the 15N HSQC of 145 μm Cdc42Δ7Q61L·GMPPNP with the indicated ratios of the TOCA1 HR1 domain, the N-WASP GBD, or both, showing that the TOCA HR1 domain does not displace the N-WASP GBD.	FIG
+163	166	HR1	structure_element	C, Selected regions of the 15N HSQC of 145 μm Cdc42Δ7Q61L·GMPPNP with the indicated ratios of the TOCA1 HR1 domain, the N-WASP GBD, or both, showing that the TOCA HR1 domain does not displace the N-WASP GBD.	FIG
+196	202	N-WASP	protein	C, Selected regions of the 15N HSQC of 145 μm Cdc42Δ7Q61L·GMPPNP with the indicated ratios of the TOCA1 HR1 domain, the N-WASP GBD, or both, showing that the TOCA HR1 domain does not displace the N-WASP GBD.	FIG
+203	206	GBD	structure_element	C, Selected regions of the 15N HSQC of 145 μm Cdc42Δ7Q61L·GMPPNP with the indicated ratios of the TOCA1 HR1 domain, the N-WASP GBD, or both, showing that the TOCA HR1 domain does not displace the N-WASP GBD.	FIG
+27	35	15N HSQC	experimental_method	D, selected regions of the 15N HSQC of 600 μm TOCA1 HR1 domain in complex with Cdc42 in the absence and presence of the N-WASP GBD, showing displacement of Cdc42 from the HR1 domain by N-WASP.	FIG
+46	51	TOCA1	protein	D, selected regions of the 15N HSQC of 600 μm TOCA1 HR1 domain in complex with Cdc42 in the absence and presence of the N-WASP GBD, showing displacement of Cdc42 from the HR1 domain by N-WASP.	FIG
+52	55	HR1	structure_element	D, selected regions of the 15N HSQC of 600 μm TOCA1 HR1 domain in complex with Cdc42 in the absence and presence of the N-WASP GBD, showing displacement of Cdc42 from the HR1 domain by N-WASP.	FIG
+63	78	in complex with	protein_state	D, selected regions of the 15N HSQC of 600 μm TOCA1 HR1 domain in complex with Cdc42 in the absence and presence of the N-WASP GBD, showing displacement of Cdc42 from the HR1 domain by N-WASP.	FIG
+79	84	Cdc42	protein	D, selected regions of the 15N HSQC of 600 μm TOCA1 HR1 domain in complex with Cdc42 in the absence and presence of the N-WASP GBD, showing displacement of Cdc42 from the HR1 domain by N-WASP.	FIG
+92	99	absence	protein_state	D, selected regions of the 15N HSQC of 600 μm TOCA1 HR1 domain in complex with Cdc42 in the absence and presence of the N-WASP GBD, showing displacement of Cdc42 from the HR1 domain by N-WASP.	FIG
+104	115	presence of	protein_state	D, selected regions of the 15N HSQC of 600 μm TOCA1 HR1 domain in complex with Cdc42 in the absence and presence of the N-WASP GBD, showing displacement of Cdc42 from the HR1 domain by N-WASP.	FIG
+120	126	N-WASP	protein	D, selected regions of the 15N HSQC of 600 μm TOCA1 HR1 domain in complex with Cdc42 in the absence and presence of the N-WASP GBD, showing displacement of Cdc42 from the HR1 domain by N-WASP.	FIG
+127	130	GBD	structure_element	D, selected regions of the 15N HSQC of 600 μm TOCA1 HR1 domain in complex with Cdc42 in the absence and presence of the N-WASP GBD, showing displacement of Cdc42 from the HR1 domain by N-WASP.	FIG
+156	161	Cdc42	protein	D, selected regions of the 15N HSQC of 600 μm TOCA1 HR1 domain in complex with Cdc42 in the absence and presence of the N-WASP GBD, showing displacement of Cdc42 from the HR1 domain by N-WASP.	FIG
+171	174	HR1	structure_element	D, selected regions of the 15N HSQC of 600 μm TOCA1 HR1 domain in complex with Cdc42 in the absence and presence of the N-WASP GBD, showing displacement of Cdc42 from the HR1 domain by N-WASP.	FIG
+185	191	N-WASP	protein	D, selected regions of the 15N HSQC of 600 μm TOCA1 HR1 domain in complex with Cdc42 in the absence and presence of the N-WASP GBD, showing displacement of Cdc42 from the HR1 domain by N-WASP.	FIG
+3	9	N-WASP	protein	An N-WASP GBD construct was produced, and its affinity for Cdc42 was measured by competition SPA (Fig. 7B).	RESULTS
+10	13	GBD	structure_element	An N-WASP GBD construct was produced, and its affinity for Cdc42 was measured by competition SPA (Fig. 7B).	RESULTS
+46	54	affinity	evidence	An N-WASP GBD construct was produced, and its affinity for Cdc42 was measured by competition SPA (Fig. 7B).	RESULTS
+59	64	Cdc42	protein	An N-WASP GBD construct was produced, and its affinity for Cdc42 was measured by competition SPA (Fig. 7B).	RESULTS
+81	96	competition SPA	experimental_method	An N-WASP GBD construct was produced, and its affinity for Cdc42 was measured by competition SPA (Fig. 7B).	RESULTS
+4	6	Kd	evidence	The Kd that was determined (37 nm) is consistent with the previously reported affinity.	RESULTS
+78	86	affinity	evidence	The Kd that was determined (37 nm) is consistent with the previously reported affinity.	RESULTS
+0	9	Unlabeled	protein_state	Unlabeled N-WASP GBD was titrated into 15N-Cdc42Δ7Q61L·GMPPNP, and the backbone NH groups were monitored using HSQCs (Fig. 7C).	RESULTS
+10	16	N-WASP	protein	Unlabeled N-WASP GBD was titrated into 15N-Cdc42Δ7Q61L·GMPPNP, and the backbone NH groups were monitored using HSQCs (Fig. 7C).	RESULTS
+17	20	GBD	structure_element	Unlabeled N-WASP GBD was titrated into 15N-Cdc42Δ7Q61L·GMPPNP, and the backbone NH groups were monitored using HSQCs (Fig. 7C).	RESULTS
+25	33	titrated	experimental_method	Unlabeled N-WASP GBD was titrated into 15N-Cdc42Δ7Q61L·GMPPNP, and the backbone NH groups were monitored using HSQCs (Fig. 7C).	RESULTS
+39	42	15N	chemical	Unlabeled N-WASP GBD was titrated into 15N-Cdc42Δ7Q61L·GMPPNP, and the backbone NH groups were monitored using HSQCs (Fig. 7C).	RESULTS
+43	61	Cdc42Δ7Q61L·GMPPNP	complex_assembly	Unlabeled N-WASP GBD was titrated into 15N-Cdc42Δ7Q61L·GMPPNP, and the backbone NH groups were monitored using HSQCs (Fig. 7C).	RESULTS
+111	116	HSQCs	experimental_method	Unlabeled N-WASP GBD was titrated into 15N-Cdc42Δ7Q61L·GMPPNP, and the backbone NH groups were monitored using HSQCs (Fig. 7C).	RESULTS
+0	9	Unlabeled	protein_state	Unlabeled HR1TOCA1 was then added to the Cdc42·N-WASP complex, and no changes were seen, suggesting that the N-WASP GBD was not displaced even in the presence of a 5-fold excess of HR1TOCA1.	RESULTS
+10	13	HR1	structure_element	Unlabeled HR1TOCA1 was then added to the Cdc42·N-WASP complex, and no changes were seen, suggesting that the N-WASP GBD was not displaced even in the presence of a 5-fold excess of HR1TOCA1.	RESULTS
+13	18	TOCA1	protein	Unlabeled HR1TOCA1 was then added to the Cdc42·N-WASP complex, and no changes were seen, suggesting that the N-WASP GBD was not displaced even in the presence of a 5-fold excess of HR1TOCA1.	RESULTS
+41	53	Cdc42·N-WASP	complex_assembly	Unlabeled HR1TOCA1 was then added to the Cdc42·N-WASP complex, and no changes were seen, suggesting that the N-WASP GBD was not displaced even in the presence of a 5-fold excess of HR1TOCA1.	RESULTS
+109	115	N-WASP	protein	Unlabeled HR1TOCA1 was then added to the Cdc42·N-WASP complex, and no changes were seen, suggesting that the N-WASP GBD was not displaced even in the presence of a 5-fold excess of HR1TOCA1.	RESULTS
+116	119	GBD	structure_element	Unlabeled HR1TOCA1 was then added to the Cdc42·N-WASP complex, and no changes were seen, suggesting that the N-WASP GBD was not displaced even in the presence of a 5-fold excess of HR1TOCA1.	RESULTS
+150	161	presence of	protein_state	Unlabeled HR1TOCA1 was then added to the Cdc42·N-WASP complex, and no changes were seen, suggesting that the N-WASP GBD was not displaced even in the presence of a 5-fold excess of HR1TOCA1.	RESULTS
+181	184	HR1	structure_element	Unlabeled HR1TOCA1 was then added to the Cdc42·N-WASP complex, and no changes were seen, suggesting that the N-WASP GBD was not displaced even in the presence of a 5-fold excess of HR1TOCA1.	RESULTS
+184	189	TOCA1	protein	Unlabeled HR1TOCA1 was then added to the Cdc42·N-WASP complex, and no changes were seen, suggesting that the N-WASP GBD was not displaced even in the presence of a 5-fold excess of HR1TOCA1.	RESULTS
+84	89	Cdc42	protein	These experiments were recorded at sufficiently high protein concentrations (145 μm Cdc42, 145 μm N-WASP GBD, 725 μm TOCA1 HR1 domain) to be far in excess of the Kd values of the individual interactions (TOCA1 Kd ≈ 5 μm, N-WASP Kd = 37 nm).	RESULTS
+98	104	N-WASP	protein	These experiments were recorded at sufficiently high protein concentrations (145 μm Cdc42, 145 μm N-WASP GBD, 725 μm TOCA1 HR1 domain) to be far in excess of the Kd values of the individual interactions (TOCA1 Kd ≈ 5 μm, N-WASP Kd = 37 nm).	RESULTS
+105	108	GBD	structure_element	These experiments were recorded at sufficiently high protein concentrations (145 μm Cdc42, 145 μm N-WASP GBD, 725 μm TOCA1 HR1 domain) to be far in excess of the Kd values of the individual interactions (TOCA1 Kd ≈ 5 μm, N-WASP Kd = 37 nm).	RESULTS
+117	122	TOCA1	protein	These experiments were recorded at sufficiently high protein concentrations (145 μm Cdc42, 145 μm N-WASP GBD, 725 μm TOCA1 HR1 domain) to be far in excess of the Kd values of the individual interactions (TOCA1 Kd ≈ 5 μm, N-WASP Kd = 37 nm).	RESULTS
+123	126	HR1	structure_element	These experiments were recorded at sufficiently high protein concentrations (145 μm Cdc42, 145 μm N-WASP GBD, 725 μm TOCA1 HR1 domain) to be far in excess of the Kd values of the individual interactions (TOCA1 Kd ≈ 5 μm, N-WASP Kd = 37 nm).	RESULTS
+162	164	Kd	evidence	These experiments were recorded at sufficiently high protein concentrations (145 μm Cdc42, 145 μm N-WASP GBD, 725 μm TOCA1 HR1 domain) to be far in excess of the Kd values of the individual interactions (TOCA1 Kd ≈ 5 μm, N-WASP Kd = 37 nm).	RESULTS
+204	209	TOCA1	protein	These experiments were recorded at sufficiently high protein concentrations (145 μm Cdc42, 145 μm N-WASP GBD, 725 μm TOCA1 HR1 domain) to be far in excess of the Kd values of the individual interactions (TOCA1 Kd ≈ 5 μm, N-WASP Kd = 37 nm).	RESULTS
+210	212	Kd	evidence	These experiments were recorded at sufficiently high protein concentrations (145 μm Cdc42, 145 μm N-WASP GBD, 725 μm TOCA1 HR1 domain) to be far in excess of the Kd values of the individual interactions (TOCA1 Kd ≈ 5 μm, N-WASP Kd = 37 nm).	RESULTS
+221	227	N-WASP	protein	These experiments were recorded at sufficiently high protein concentrations (145 μm Cdc42, 145 μm N-WASP GBD, 725 μm TOCA1 HR1 domain) to be far in excess of the Kd values of the individual interactions (TOCA1 Kd ≈ 5 μm, N-WASP Kd = 37 nm).	RESULTS
+228	230	Kd	evidence	These experiments were recorded at sufficiently high protein concentrations (145 μm Cdc42, 145 μm N-WASP GBD, 725 μm TOCA1 HR1 domain) to be far in excess of the Kd values of the individual interactions (TOCA1 Kd ≈ 5 μm, N-WASP Kd = 37 nm).	RESULTS
+20	24	HSQC	experimental_method	A comparison of the HSQC experiments recorded on 15N-Cdc42 alone, in the presence of TOCA1 HR1, N-WASP GBD, or both, shows that the spectra in the presence of N-WASP and in the presence of both N-WASP and TOCA1 HR1 are identical (Fig. 7C).	RESULTS
+49	52	15N	chemical	A comparison of the HSQC experiments recorded on 15N-Cdc42 alone, in the presence of TOCA1 HR1, N-WASP GBD, or both, shows that the spectra in the presence of N-WASP and in the presence of both N-WASP and TOCA1 HR1 are identical (Fig. 7C).	RESULTS
+53	58	Cdc42	protein	A comparison of the HSQC experiments recorded on 15N-Cdc42 alone, in the presence of TOCA1 HR1, N-WASP GBD, or both, shows that the spectra in the presence of N-WASP and in the presence of both N-WASP and TOCA1 HR1 are identical (Fig. 7C).	RESULTS
+59	64	alone	protein_state	A comparison of the HSQC experiments recorded on 15N-Cdc42 alone, in the presence of TOCA1 HR1, N-WASP GBD, or both, shows that the spectra in the presence of N-WASP and in the presence of both N-WASP and TOCA1 HR1 are identical (Fig. 7C).	RESULTS
+73	84	presence of	protein_state	A comparison of the HSQC experiments recorded on 15N-Cdc42 alone, in the presence of TOCA1 HR1, N-WASP GBD, or both, shows that the spectra in the presence of N-WASP and in the presence of both N-WASP and TOCA1 HR1 are identical (Fig. 7C).	RESULTS
+85	90	TOCA1	protein	A comparison of the HSQC experiments recorded on 15N-Cdc42 alone, in the presence of TOCA1 HR1, N-WASP GBD, or both, shows that the spectra in the presence of N-WASP and in the presence of both N-WASP and TOCA1 HR1 are identical (Fig. 7C).	RESULTS
+91	94	HR1	structure_element	A comparison of the HSQC experiments recorded on 15N-Cdc42 alone, in the presence of TOCA1 HR1, N-WASP GBD, or both, shows that the spectra in the presence of N-WASP and in the presence of both N-WASP and TOCA1 HR1 are identical (Fig. 7C).	RESULTS
+96	102	N-WASP	protein	A comparison of the HSQC experiments recorded on 15N-Cdc42 alone, in the presence of TOCA1 HR1, N-WASP GBD, or both, shows that the spectra in the presence of N-WASP and in the presence of both N-WASP and TOCA1 HR1 are identical (Fig. 7C).	RESULTS
+103	106	GBD	structure_element	A comparison of the HSQC experiments recorded on 15N-Cdc42 alone, in the presence of TOCA1 HR1, N-WASP GBD, or both, shows that the spectra in the presence of N-WASP and in the presence of both N-WASP and TOCA1 HR1 are identical (Fig. 7C).	RESULTS
+132	139	spectra	evidence	A comparison of the HSQC experiments recorded on 15N-Cdc42 alone, in the presence of TOCA1 HR1, N-WASP GBD, or both, shows that the spectra in the presence of N-WASP and in the presence of both N-WASP and TOCA1 HR1 are identical (Fig. 7C).	RESULTS
+147	158	presence of	protein_state	A comparison of the HSQC experiments recorded on 15N-Cdc42 alone, in the presence of TOCA1 HR1, N-WASP GBD, or both, shows that the spectra in the presence of N-WASP and in the presence of both N-WASP and TOCA1 HR1 are identical (Fig. 7C).	RESULTS
+159	165	N-WASP	protein	A comparison of the HSQC experiments recorded on 15N-Cdc42 alone, in the presence of TOCA1 HR1, N-WASP GBD, or both, shows that the spectra in the presence of N-WASP and in the presence of both N-WASP and TOCA1 HR1 are identical (Fig. 7C).	RESULTS
+177	188	presence of	protein_state	A comparison of the HSQC experiments recorded on 15N-Cdc42 alone, in the presence of TOCA1 HR1, N-WASP GBD, or both, shows that the spectra in the presence of N-WASP and in the presence of both N-WASP and TOCA1 HR1 are identical (Fig. 7C).	RESULTS
+194	200	N-WASP	protein	A comparison of the HSQC experiments recorded on 15N-Cdc42 alone, in the presence of TOCA1 HR1, N-WASP GBD, or both, shows that the spectra in the presence of N-WASP and in the presence of both N-WASP and TOCA1 HR1 are identical (Fig. 7C).	RESULTS
+205	210	TOCA1	protein	A comparison of the HSQC experiments recorded on 15N-Cdc42 alone, in the presence of TOCA1 HR1, N-WASP GBD, or both, shows that the spectra in the presence of N-WASP and in the presence of both N-WASP and TOCA1 HR1 are identical (Fig. 7C).	RESULTS
+211	214	HR1	structure_element	A comparison of the HSQC experiments recorded on 15N-Cdc42 alone, in the presence of TOCA1 HR1, N-WASP GBD, or both, shows that the spectra in the presence of N-WASP and in the presence of both N-WASP and TOCA1 HR1 are identical (Fig. 7C).	RESULTS
+13	16	15N	chemical	Furthermore, 15N-TOCA1 HR1 was monitored in the presence of unlabeled Cdc42Δ7Q61L·GMPPNP (1:1) before and after the addition of 0.25 and 1.0 eq of unlabeled N-WASP GBD.	RESULTS
+17	22	TOCA1	protein	Furthermore, 15N-TOCA1 HR1 was monitored in the presence of unlabeled Cdc42Δ7Q61L·GMPPNP (1:1) before and after the addition of 0.25 and 1.0 eq of unlabeled N-WASP GBD.	RESULTS
+23	26	HR1	structure_element	Furthermore, 15N-TOCA1 HR1 was monitored in the presence of unlabeled Cdc42Δ7Q61L·GMPPNP (1:1) before and after the addition of 0.25 and 1.0 eq of unlabeled N-WASP GBD.	RESULTS
+48	59	presence of	protein_state	Furthermore, 15N-TOCA1 HR1 was monitored in the presence of unlabeled Cdc42Δ7Q61L·GMPPNP (1:1) before and after the addition of 0.25 and 1.0 eq of unlabeled N-WASP GBD.	RESULTS
+60	69	unlabeled	protein_state	Furthermore, 15N-TOCA1 HR1 was monitored in the presence of unlabeled Cdc42Δ7Q61L·GMPPNP (1:1) before and after the addition of 0.25 and 1.0 eq of unlabeled N-WASP GBD.	RESULTS
+70	88	Cdc42Δ7Q61L·GMPPNP	complex_assembly	Furthermore, 15N-TOCA1 HR1 was monitored in the presence of unlabeled Cdc42Δ7Q61L·GMPPNP (1:1) before and after the addition of 0.25 and 1.0 eq of unlabeled N-WASP GBD.	RESULTS
+147	156	unlabeled	protein_state	Furthermore, 15N-TOCA1 HR1 was monitored in the presence of unlabeled Cdc42Δ7Q61L·GMPPNP (1:1) before and after the addition of 0.25 and 1.0 eq of unlabeled N-WASP GBD.	RESULTS
+157	163	N-WASP	protein	Furthermore, 15N-TOCA1 HR1 was monitored in the presence of unlabeled Cdc42Δ7Q61L·GMPPNP (1:1) before and after the addition of 0.25 and 1.0 eq of unlabeled N-WASP GBD.	RESULTS
+164	167	GBD	structure_element	Furthermore, 15N-TOCA1 HR1 was monitored in the presence of unlabeled Cdc42Δ7Q61L·GMPPNP (1:1) before and after the addition of 0.25 and 1.0 eq of unlabeled N-WASP GBD.	RESULTS
+4	12	spectrum	evidence	The spectrum when N-WASP and TOCA1 were equimolar was identical to that of the free HR1 domain, whereas the spectrum in the presence of 0.25 eq of N-WASP was intermediate between the TOCA1 HR1 free and complex spectra (Fig. 7D).	RESULTS
+18	24	N-WASP	protein	The spectrum when N-WASP and TOCA1 were equimolar was identical to that of the free HR1 domain, whereas the spectrum in the presence of 0.25 eq of N-WASP was intermediate between the TOCA1 HR1 free and complex spectra (Fig. 7D).	RESULTS
+29	34	TOCA1	protein	The spectrum when N-WASP and TOCA1 were equimolar was identical to that of the free HR1 domain, whereas the spectrum in the presence of 0.25 eq of N-WASP was intermediate between the TOCA1 HR1 free and complex spectra (Fig. 7D).	RESULTS
+79	83	free	protein_state	The spectrum when N-WASP and TOCA1 were equimolar was identical to that of the free HR1 domain, whereas the spectrum in the presence of 0.25 eq of N-WASP was intermediate between the TOCA1 HR1 free and complex spectra (Fig. 7D).	RESULTS
+84	87	HR1	structure_element	The spectrum when N-WASP and TOCA1 were equimolar was identical to that of the free HR1 domain, whereas the spectrum in the presence of 0.25 eq of N-WASP was intermediate between the TOCA1 HR1 free and complex spectra (Fig. 7D).	RESULTS
+108	116	spectrum	evidence	The spectrum when N-WASP and TOCA1 were equimolar was identical to that of the free HR1 domain, whereas the spectrum in the presence of 0.25 eq of N-WASP was intermediate between the TOCA1 HR1 free and complex spectra (Fig. 7D).	RESULTS
+124	135	presence of	protein_state	The spectrum when N-WASP and TOCA1 were equimolar was identical to that of the free HR1 domain, whereas the spectrum in the presence of 0.25 eq of N-WASP was intermediate between the TOCA1 HR1 free and complex spectra (Fig. 7D).	RESULTS
+147	153	N-WASP	protein	The spectrum when N-WASP and TOCA1 were equimolar was identical to that of the free HR1 domain, whereas the spectrum in the presence of 0.25 eq of N-WASP was intermediate between the TOCA1 HR1 free and complex spectra (Fig. 7D).	RESULTS
+183	188	TOCA1	protein	The spectrum when N-WASP and TOCA1 were equimolar was identical to that of the free HR1 domain, whereas the spectrum in the presence of 0.25 eq of N-WASP was intermediate between the TOCA1 HR1 free and complex spectra (Fig. 7D).	RESULTS
+189	192	HR1	structure_element	The spectrum when N-WASP and TOCA1 were equimolar was identical to that of the free HR1 domain, whereas the spectrum in the presence of 0.25 eq of N-WASP was intermediate between the TOCA1 HR1 free and complex spectra (Fig. 7D).	RESULTS
+193	197	free	protein_state	The spectrum when N-WASP and TOCA1 were equimolar was identical to that of the free HR1 domain, whereas the spectrum in the presence of 0.25 eq of N-WASP was intermediate between the TOCA1 HR1 free and complex spectra (Fig. 7D).	RESULTS
+202	209	complex	protein_state	The spectrum when N-WASP and TOCA1 were equimolar was identical to that of the free HR1 domain, whereas the spectrum in the presence of 0.25 eq of N-WASP was intermediate between the TOCA1 HR1 free and complex spectra (Fig. 7D).	RESULTS
+210	217	spectra	evidence	The spectrum when N-WASP and TOCA1 were equimolar was identical to that of the free HR1 domain, whereas the spectrum in the presence of 0.25 eq of N-WASP was intermediate between the TOCA1 HR1 free and complex spectra (Fig. 7D).	RESULTS
+27	30	NMR	experimental_method	When in fast exchange, the NMR signal represents a population-weighted average between free and bound states, so the intermediate spectrum indicates that the population comprises a mixture of free and bound HR1 domain.	RESULTS
+87	91	free	protein_state	When in fast exchange, the NMR signal represents a population-weighted average between free and bound states, so the intermediate spectrum indicates that the population comprises a mixture of free and bound HR1 domain.	RESULTS
+96	101	bound	protein_state	When in fast exchange, the NMR signal represents a population-weighted average between free and bound states, so the intermediate spectrum indicates that the population comprises a mixture of free and bound HR1 domain.	RESULTS
+130	138	spectrum	evidence	When in fast exchange, the NMR signal represents a population-weighted average between free and bound states, so the intermediate spectrum indicates that the population comprises a mixture of free and bound HR1 domain.	RESULTS
+192	196	free	protein_state	When in fast exchange, the NMR signal represents a population-weighted average between free and bound states, so the intermediate spectrum indicates that the population comprises a mixture of free and bound HR1 domain.	RESULTS
+201	206	bound	protein_state	When in fast exchange, the NMR signal represents a population-weighted average between free and bound states, so the intermediate spectrum indicates that the population comprises a mixture of free and bound HR1 domain.	RESULTS
+207	210	HR1	structure_element	When in fast exchange, the NMR signal represents a population-weighted average between free and bound states, so the intermediate spectrum indicates that the population comprises a mixture of free and bound HR1 domain.	RESULTS
+88	90	Kd	evidence	Again, the experiments were recorded on protein samples far in excess of the individual Kd values (600 μm each protein).	RESULTS
+29	32	HR1	structure_element	These data indicate that the HR1 domain is displaced from Cdc42 by N-WASP and that a ternary complex comprising TOCA1 HR1, N-WASP GBD, and Cdc42 is not formed.	RESULTS
+58	63	Cdc42	protein	These data indicate that the HR1 domain is displaced from Cdc42 by N-WASP and that a ternary complex comprising TOCA1 HR1, N-WASP GBD, and Cdc42 is not formed.	RESULTS
+67	73	N-WASP	protein	These data indicate that the HR1 domain is displaced from Cdc42 by N-WASP and that a ternary complex comprising TOCA1 HR1, N-WASP GBD, and Cdc42 is not formed.	RESULTS
+112	117	TOCA1	protein	These data indicate that the HR1 domain is displaced from Cdc42 by N-WASP and that a ternary complex comprising TOCA1 HR1, N-WASP GBD, and Cdc42 is not formed.	RESULTS
+118	121	HR1	structure_element	These data indicate that the HR1 domain is displaced from Cdc42 by N-WASP and that a ternary complex comprising TOCA1 HR1, N-WASP GBD, and Cdc42 is not formed.	RESULTS
+123	129	N-WASP	protein	These data indicate that the HR1 domain is displaced from Cdc42 by N-WASP and that a ternary complex comprising TOCA1 HR1, N-WASP GBD, and Cdc42 is not formed.	RESULTS
+130	133	GBD	structure_element	These data indicate that the HR1 domain is displaced from Cdc42 by N-WASP and that a ternary complex comprising TOCA1 HR1, N-WASP GBD, and Cdc42 is not formed.	RESULTS
+139	144	Cdc42	protein	These data indicate that the HR1 domain is displaced from Cdc42 by N-WASP and that a ternary complex comprising TOCA1 HR1, N-WASP GBD, and Cdc42 is not formed.	RESULTS
+85	90	Cdc42	protein	Taken together, the data in Fig. 7, C and D, indicate unidirectional competition for Cdc42 binding in which the N-WASP GBD displaces TOCA1 HR1 but not vice versa.	RESULTS
+112	118	N-WASP	protein	Taken together, the data in Fig. 7, C and D, indicate unidirectional competition for Cdc42 binding in which the N-WASP GBD displaces TOCA1 HR1 but not vice versa.	RESULTS
+119	122	GBD	structure_element	Taken together, the data in Fig. 7, C and D, indicate unidirectional competition for Cdc42 binding in which the N-WASP GBD displaces TOCA1 HR1 but not vice versa.	RESULTS
+133	138	TOCA1	protein	Taken together, the data in Fig. 7, C and D, indicate unidirectional competition for Cdc42 binding in which the N-WASP GBD displaces TOCA1 HR1 but not vice versa.	RESULTS
+139	142	HR1	structure_element	Taken together, the data in Fig. 7, C and D, indicate unidirectional competition for Cdc42 binding in which the N-WASP GBD displaces TOCA1 HR1 but not vice versa.	RESULTS
+78	83	TOCA1	protein	To extend these studies to a more complex system and to assess the ability of TOCA1 HR1 to compete with full-length N-WASP, pyrene actin assays were employed.	RESULTS
+84	87	HR1	structure_element	To extend these studies to a more complex system and to assess the ability of TOCA1 HR1 to compete with full-length N-WASP, pyrene actin assays were employed.	RESULTS
+104	115	full-length	protein_state	To extend these studies to a more complex system and to assess the ability of TOCA1 HR1 to compete with full-length N-WASP, pyrene actin assays were employed.	RESULTS
+116	122	N-WASP	protein	To extend these studies to a more complex system and to assess the ability of TOCA1 HR1 to compete with full-length N-WASP, pyrene actin assays were employed.	RESULTS
+124	143	pyrene actin assays	experimental_method	To extend these studies to a more complex system and to assess the ability of TOCA1 HR1 to compete with full-length N-WASP, pyrene actin assays were employed.	RESULTS
+68	80	pyrene actin	chemical	These assays, described in detail elsewhere, were carried out using pyrene actin-supplemented Xenopus extracts into which exogenous TOCA1 HR1 domain or N-WASP GBD was added, to assess their effects on actin polymerization.	RESULTS
+94	101	Xenopus	taxonomy_domain	These assays, described in detail elsewhere, were carried out using pyrene actin-supplemented Xenopus extracts into which exogenous TOCA1 HR1 domain or N-WASP GBD was added, to assess their effects on actin polymerization.	RESULTS
+132	137	TOCA1	protein	These assays, described in detail elsewhere, were carried out using pyrene actin-supplemented Xenopus extracts into which exogenous TOCA1 HR1 domain or N-WASP GBD was added, to assess their effects on actin polymerization.	RESULTS
+138	141	HR1	structure_element	These assays, described in detail elsewhere, were carried out using pyrene actin-supplemented Xenopus extracts into which exogenous TOCA1 HR1 domain or N-WASP GBD was added, to assess their effects on actin polymerization.	RESULTS
+152	158	N-WASP	protein	These assays, described in detail elsewhere, were carried out using pyrene actin-supplemented Xenopus extracts into which exogenous TOCA1 HR1 domain or N-WASP GBD was added, to assess their effects on actin polymerization.	RESULTS
+159	162	GBD	structure_element	These assays, described in detail elsewhere, were carried out using pyrene actin-supplemented Xenopus extracts into which exogenous TOCA1 HR1 domain or N-WASP GBD was added, to assess their effects on actin polymerization.	RESULTS
+201	206	actin	protein_type	These assays, described in detail elsewhere, were carried out using pyrene actin-supplemented Xenopus extracts into which exogenous TOCA1 HR1 domain or N-WASP GBD was added, to assess their effects on actin polymerization.	RESULTS
+0	5	Actin	protein_type	Actin polymerization in all cases was initiated by the addition of PI(4,5)P2-containing liposomes.	RESULTS
+67	76	PI(4,5)P2	chemical	Actin polymerization in all cases was initiated by the addition of PI(4,5)P2-containing liposomes.	RESULTS
+0	5	Actin	protein_type	Actin polymerization triggered by the addition of PI(4,5)P2-containing liposomes has previously been shown to depend on TOCA1 and N-WASP.	RESULTS
+50	59	PI(4,5)P2	chemical	Actin polymerization triggered by the addition of PI(4,5)P2-containing liposomes has previously been shown to depend on TOCA1 and N-WASP.	RESULTS
+120	125	TOCA1	protein	Actin polymerization triggered by the addition of PI(4,5)P2-containing liposomes has previously been shown to depend on TOCA1 and N-WASP.	RESULTS
+130	136	N-WASP	protein	Actin polymerization triggered by the addition of PI(4,5)P2-containing liposomes has previously been shown to depend on TOCA1 and N-WASP.	RESULTS
+11	17	N-WASP	protein	Endogenous N-WASP is present at ∼100 nm in Xenopus extracts, whereas TOCA1 is present at a 10-fold lower concentration than N-WASP.	RESULTS
+43	50	Xenopus	taxonomy_domain	Endogenous N-WASP is present at ∼100 nm in Xenopus extracts, whereas TOCA1 is present at a 10-fold lower concentration than N-WASP.	RESULTS
+69	74	TOCA1	protein	Endogenous N-WASP is present at ∼100 nm in Xenopus extracts, whereas TOCA1 is present at a 10-fold lower concentration than N-WASP.	RESULTS
+124	130	N-WASP	protein	Endogenous N-WASP is present at ∼100 nm in Xenopus extracts, whereas TOCA1 is present at a 10-fold lower concentration than N-WASP.	RESULTS
+4	12	addition	experimental_method	The addition of the isolated N-WASP GBD significantly inhibited the polymerization of actin at concentrations as low as 100 nm and completely abolished polymerization at higher concentrations (Fig. 8).	RESULTS
+29	35	N-WASP	protein	The addition of the isolated N-WASP GBD significantly inhibited the polymerization of actin at concentrations as low as 100 nm and completely abolished polymerization at higher concentrations (Fig. 8).	RESULTS
+36	39	GBD	structure_element	The addition of the isolated N-WASP GBD significantly inhibited the polymerization of actin at concentrations as low as 100 nm and completely abolished polymerization at higher concentrations (Fig. 8).	RESULTS
+86	91	actin	protein_type	The addition of the isolated N-WASP GBD significantly inhibited the polymerization of actin at concentrations as low as 100 nm and completely abolished polymerization at higher concentrations (Fig. 8).	RESULTS
+4	7	GBD	structure_element	The GBD presumably acts as a dominant negative, sequestering endogenous Cdc42 and preventing endogenous full-length N-WASP from binding and becoming activated.	RESULTS
+72	77	Cdc42	protein	The GBD presumably acts as a dominant negative, sequestering endogenous Cdc42 and preventing endogenous full-length N-WASP from binding and becoming activated.	RESULTS
+93	103	endogenous	protein_state	The GBD presumably acts as a dominant negative, sequestering endogenous Cdc42 and preventing endogenous full-length N-WASP from binding and becoming activated.	RESULTS
+104	115	full-length	protein_state	The GBD presumably acts as a dominant negative, sequestering endogenous Cdc42 and preventing endogenous full-length N-WASP from binding and becoming activated.	RESULTS
+116	122	N-WASP	protein	The GBD presumably acts as a dominant negative, sequestering endogenous Cdc42 and preventing endogenous full-length N-WASP from binding and becoming activated.	RESULTS
+4	12	addition	experimental_method	The addition of the TOCA1 HR1 domain to 100 μm had no significant effect on the rate of actin polymerization or maximum fluorescence.	RESULTS
+20	25	TOCA1	protein	The addition of the TOCA1 HR1 domain to 100 μm had no significant effect on the rate of actin polymerization or maximum fluorescence.	RESULTS
+26	29	HR1	structure_element	The addition of the TOCA1 HR1 domain to 100 μm had no significant effect on the rate of actin polymerization or maximum fluorescence.	RESULTS
+88	93	actin	protein_type	The addition of the TOCA1 HR1 domain to 100 μm had no significant effect on the rate of actin polymerization or maximum fluorescence.	RESULTS
+112	132	maximum fluorescence	evidence	The addition of the TOCA1 HR1 domain to 100 μm had no significant effect on the rate of actin polymerization or maximum fluorescence.	RESULTS
+24	34	endogenous	protein_state	This is consistent with endogenous N-WASP, activated by other components of the assay, outcompeting the TOCA1 HR1 domain for Cdc42 binding.	RESULTS
+35	41	N-WASP	protein	This is consistent with endogenous N-WASP, activated by other components of the assay, outcompeting the TOCA1 HR1 domain for Cdc42 binding.	RESULTS
+104	109	TOCA1	protein	This is consistent with endogenous N-WASP, activated by other components of the assay, outcompeting the TOCA1 HR1 domain for Cdc42 binding.	RESULTS
+110	113	HR1	structure_element	This is consistent with endogenous N-WASP, activated by other components of the assay, outcompeting the TOCA1 HR1 domain for Cdc42 binding.	RESULTS
+125	130	Cdc42	protein	This is consistent with endogenous N-WASP, activated by other components of the assay, outcompeting the TOCA1 HR1 domain for Cdc42 binding.	RESULTS
+35	53	Cdc42·N-WASP·TOCA1	complex_assembly	Actin polymerization downstream of Cdc42·N-WASP·TOCA1 is inhibited by excess N-WASP GBD but not by the TOCA1 HR1 domain.	FIG
+57	66	inhibited	protein_state	Actin polymerization downstream of Cdc42·N-WASP·TOCA1 is inhibited by excess N-WASP GBD but not by the TOCA1 HR1 domain.	FIG
+77	83	N-WASP	protein	Actin polymerization downstream of Cdc42·N-WASP·TOCA1 is inhibited by excess N-WASP GBD but not by the TOCA1 HR1 domain.	FIG
+84	87	GBD	structure_element	Actin polymerization downstream of Cdc42·N-WASP·TOCA1 is inhibited by excess N-WASP GBD but not by the TOCA1 HR1 domain.	FIG
+103	108	TOCA1	protein	Actin polymerization downstream of Cdc42·N-WASP·TOCA1 is inhibited by excess N-WASP GBD but not by the TOCA1 HR1 domain.	FIG
+109	112	HR1	structure_element	Actin polymerization downstream of Cdc42·N-WASP·TOCA1 is inhibited by excess N-WASP GBD but not by the TOCA1 HR1 domain.	FIG
+0	19	Fluorescence curves	evidence	Fluorescence curves show actin polymerization in the presence of increasing concentrations of N-WASP GBD or TOCA1 HR1 domain as indicated.	FIG
+53	64	presence of	protein_state	Fluorescence curves show actin polymerization in the presence of increasing concentrations of N-WASP GBD or TOCA1 HR1 domain as indicated.	FIG
+65	90	increasing concentrations	experimental_method	Fluorescence curves show actin polymerization in the presence of increasing concentrations of N-WASP GBD or TOCA1 HR1 domain as indicated.	FIG
+94	100	N-WASP	protein	Fluorescence curves show actin polymerization in the presence of increasing concentrations of N-WASP GBD or TOCA1 HR1 domain as indicated.	FIG
+101	104	GBD	structure_element	Fluorescence curves show actin polymerization in the presence of increasing concentrations of N-WASP GBD or TOCA1 HR1 domain as indicated.	FIG
+108	113	TOCA1	protein	Fluorescence curves show actin polymerization in the presence of increasing concentrations of N-WASP GBD or TOCA1 HR1 domain as indicated.	FIG
+114	117	HR1	structure_element	Fluorescence curves show actin polymerization in the presence of increasing concentrations of N-WASP GBD or TOCA1 HR1 domain as indicated.	FIG
+4	9	Cdc42	protein	The Cdc42-TOCA1 Interaction	DISCUSS
+10	15	TOCA1	protein	The Cdc42-TOCA1 Interaction	DISCUSS
+4	9	TOCA1	protein	The TOCA1 HR1 domain alone is sufficient for Cdc42 binding in vitro, yet the affinity of the TOCA1 HR1 domain for Cdc42 is remarkably low (Kd ≈ 5 μm).	DISCUSS
+10	13	HR1	structure_element	The TOCA1 HR1 domain alone is sufficient for Cdc42 binding in vitro, yet the affinity of the TOCA1 HR1 domain for Cdc42 is remarkably low (Kd ≈ 5 μm).	DISCUSS
+21	26	alone	protein_state	The TOCA1 HR1 domain alone is sufficient for Cdc42 binding in vitro, yet the affinity of the TOCA1 HR1 domain for Cdc42 is remarkably low (Kd ≈ 5 μm).	DISCUSS
+45	50	Cdc42	protein	The TOCA1 HR1 domain alone is sufficient for Cdc42 binding in vitro, yet the affinity of the TOCA1 HR1 domain for Cdc42 is remarkably low (Kd ≈ 5 μm).	DISCUSS
+77	85	affinity	evidence	The TOCA1 HR1 domain alone is sufficient for Cdc42 binding in vitro, yet the affinity of the TOCA1 HR1 domain for Cdc42 is remarkably low (Kd ≈ 5 μm).	DISCUSS
+93	98	TOCA1	protein	The TOCA1 HR1 domain alone is sufficient for Cdc42 binding in vitro, yet the affinity of the TOCA1 HR1 domain for Cdc42 is remarkably low (Kd ≈ 5 μm).	DISCUSS
+99	102	HR1	structure_element	The TOCA1 HR1 domain alone is sufficient for Cdc42 binding in vitro, yet the affinity of the TOCA1 HR1 domain for Cdc42 is remarkably low (Kd ≈ 5 μm).	DISCUSS
+114	119	Cdc42	protein	The TOCA1 HR1 domain alone is sufficient for Cdc42 binding in vitro, yet the affinity of the TOCA1 HR1 domain for Cdc42 is remarkably low (Kd ≈ 5 μm).	DISCUSS
+139	141	Kd	evidence	The TOCA1 HR1 domain alone is sufficient for Cdc42 binding in vitro, yet the affinity of the TOCA1 HR1 domain for Cdc42 is remarkably low (Kd ≈ 5 μm).	DISCUSS
+46	52	N-WASP	protein	This is over 100 times lower than that of the N-WASP GBD (Kd = 37 nm) and considerably lower than other known G protein-HR1 domain interactions.	DISCUSS
+53	56	GBD	structure_element	This is over 100 times lower than that of the N-WASP GBD (Kd = 37 nm) and considerably lower than other known G protein-HR1 domain interactions.	DISCUSS
+58	60	Kd	evidence	This is over 100 times lower than that of the N-WASP GBD (Kd = 37 nm) and considerably lower than other known G protein-HR1 domain interactions.	DISCUSS
+110	119	G protein	protein_type	This is over 100 times lower than that of the N-WASP GBD (Kd = 37 nm) and considerably lower than other known G protein-HR1 domain interactions.	DISCUSS
+120	123	HR1	structure_element	This is over 100 times lower than that of the N-WASP GBD (Kd = 37 nm) and considerably lower than other known G protein-HR1 domain interactions.	DISCUSS
+31	48	C-terminal region	structure_element	The polybasic tract within the C-terminal region of Cdc42 does not appear to be required for binding to TOCA1, which is in contrast to the interaction between Rac1 and the HR1b domain of PRK1 but more similar to the PRK1 HR1a-RhoA interaction.	DISCUSS
+52	57	Cdc42	protein	The polybasic tract within the C-terminal region of Cdc42 does not appear to be required for binding to TOCA1, which is in contrast to the interaction between Rac1 and the HR1b domain of PRK1 but more similar to the PRK1 HR1a-RhoA interaction.	DISCUSS
+104	109	TOCA1	protein	The polybasic tract within the C-terminal region of Cdc42 does not appear to be required for binding to TOCA1, which is in contrast to the interaction between Rac1 and the HR1b domain of PRK1 but more similar to the PRK1 HR1a-RhoA interaction.	DISCUSS
+159	163	Rac1	protein	The polybasic tract within the C-terminal region of Cdc42 does not appear to be required for binding to TOCA1, which is in contrast to the interaction between Rac1 and the HR1b domain of PRK1 but more similar to the PRK1 HR1a-RhoA interaction.	DISCUSS
+172	176	HR1b	structure_element	The polybasic tract within the C-terminal region of Cdc42 does not appear to be required for binding to TOCA1, which is in contrast to the interaction between Rac1 and the HR1b domain of PRK1 but more similar to the PRK1 HR1a-RhoA interaction.	DISCUSS
+187	191	PRK1	protein	The polybasic tract within the C-terminal region of Cdc42 does not appear to be required for binding to TOCA1, which is in contrast to the interaction between Rac1 and the HR1b domain of PRK1 but more similar to the PRK1 HR1a-RhoA interaction.	DISCUSS
+216	220	PRK1	protein	The polybasic tract within the C-terminal region of Cdc42 does not appear to be required for binding to TOCA1, which is in contrast to the interaction between Rac1 and the HR1b domain of PRK1 but more similar to the PRK1 HR1a-RhoA interaction.	DISCUSS
+221	225	HR1a	structure_element	The polybasic tract within the C-terminal region of Cdc42 does not appear to be required for binding to TOCA1, which is in contrast to the interaction between Rac1 and the HR1b domain of PRK1 but more similar to the PRK1 HR1a-RhoA interaction.	DISCUSS
+226	230	RhoA	protein	The polybasic tract within the C-terminal region of Cdc42 does not appear to be required for binding to TOCA1, which is in contrast to the interaction between Rac1 and the HR1b domain of PRK1 but more similar to the PRK1 HR1a-RhoA interaction.	DISCUSS
+9	26	binding interface	site	A single binding interface on both the HR1 domain and Cdc42 can be concluded from the data presented here.	DISCUSS
+39	42	HR1	structure_element	A single binding interface on both the HR1 domain and Cdc42 can be concluded from the data presented here.	DISCUSS
+54	59	Cdc42	protein	A single binding interface on both the HR1 domain and Cdc42 can be concluded from the data presented here.	DISCUSS
+17	27	interfaces	site	Furthermore, the interfaces are comparable with those of other G protein-HR1 interactions (Fig. 4), and the lowest energy model produced in rigid body docking resembles previously studied G protein·HR1 complexes (Fig. 6).	DISCUSS
+63	72	G protein	protein_type	Furthermore, the interfaces are comparable with those of other G protein-HR1 interactions (Fig. 4), and the lowest energy model produced in rigid body docking resembles previously studied G protein·HR1 complexes (Fig. 6).	DISCUSS
+73	76	HR1	structure_element	Furthermore, the interfaces are comparable with those of other G protein-HR1 interactions (Fig. 4), and the lowest energy model produced in rigid body docking resembles previously studied G protein·HR1 complexes (Fig. 6).	DISCUSS
+122	127	model	evidence	Furthermore, the interfaces are comparable with those of other G protein-HR1 interactions (Fig. 4), and the lowest energy model produced in rigid body docking resembles previously studied G protein·HR1 complexes (Fig. 6).	DISCUSS
+140	158	rigid body docking	experimental_method	Furthermore, the interfaces are comparable with those of other G protein-HR1 interactions (Fig. 4), and the lowest energy model produced in rigid body docking resembles previously studied G protein·HR1 complexes (Fig. 6).	DISCUSS
+188	201	G protein·HR1	complex_assembly	Furthermore, the interfaces are comparable with those of other G protein-HR1 interactions (Fig. 4), and the lowest energy model produced in rigid body docking resembles previously studied G protein·HR1 complexes (Fig. 6).	DISCUSS
+124	127	HR1	structure_element	It seems, therefore, that the interaction, despite its relatively low affinity, is specific and sterically similar to other HR1 domain-G protein interactions.	DISCUSS
+135	144	G protein	protein_type	It seems, therefore, that the interaction, despite its relatively low affinity, is specific and sterically similar to other HR1 domain-G protein interactions.	DISCUSS
+4	9	TOCA1	protein	The TOCA1 HR1 domain is a left-handed coiled-coil comparable with other known HR1 domains.	DISCUSS
+10	13	HR1	structure_element	The TOCA1 HR1 domain is a left-handed coiled-coil comparable with other known HR1 domains.	DISCUSS
+38	49	coiled-coil	structure_element	The TOCA1 HR1 domain is a left-handed coiled-coil comparable with other known HR1 domains.	DISCUSS
+78	81	HR1	structure_element	The TOCA1 HR1 domain is a left-handed coiled-coil comparable with other known HR1 domains.	DISCUSS
+33	44	coiled-coil	structure_element	A short region N-terminal to the coiled-coil exhibits a series of turns and contacts residues of both helices of the coiled-coil (Fig. 3).	DISCUSS
+117	128	coiled-coil	structure_element	A short region N-terminal to the coiled-coil exhibits a series of turns and contacts residues of both helices of the coiled-coil (Fig. 3).	DISCUSS
+30	34	CIP4	protein	The corresponding sequence in CIP4 also includes a series of turns but is flexible, whereas in the HR1a domain of PRK1, the equivalent region adopts an α-helical structure that packs against the coiled-coil.	DISCUSS
+99	103	HR1a	structure_element	The corresponding sequence in CIP4 also includes a series of turns but is flexible, whereas in the HR1a domain of PRK1, the equivalent region adopts an α-helical structure that packs against the coiled-coil.	DISCUSS
+114	118	PRK1	protein	The corresponding sequence in CIP4 also includes a series of turns but is flexible, whereas in the HR1a domain of PRK1, the equivalent region adopts an α-helical structure that packs against the coiled-coil.	DISCUSS
+152	171	α-helical structure	structure_element	The corresponding sequence in CIP4 also includes a series of turns but is flexible, whereas in the HR1a domain of PRK1, the equivalent region adopts an α-helical structure that packs against the coiled-coil.	DISCUSS
+195	206	coiled-coil	structure_element	The corresponding sequence in CIP4 also includes a series of turns but is flexible, whereas in the HR1a domain of PRK1, the equivalent region adopts an α-helical structure that packs against the coiled-coil.	DISCUSS
+51	62	coiled-coil	structure_element	The contacts between the N-terminal region and the coiled-coil are predominantly hydrophobic in both cases, but sequence-specific contacts do not appear to be conserved.	DISCUSS
+32	59	G protein-binding interface	site	This region is distant from the G protein-binding interface of the HR1 domains, so the structural differences may relate to the structure and regulation of these domains rather than their G protein interactions.	DISCUSS
+67	70	HR1	structure_element	This region is distant from the G protein-binding interface of the HR1 domains, so the structural differences may relate to the structure and regulation of these domains rather than their G protein interactions.	DISCUSS
+188	197	G protein	protein_type	This region is distant from the G protein-binding interface of the HR1 domains, so the structural differences may relate to the structure and regulation of these domains rather than their G protein interactions.	DISCUSS
+4	22	interhelical loops	structure_element	The interhelical loops of TOCA1 and CIP4 differ from the same region in the HR1 domains of PRK1 in that they are longer and contain two short stretches of 310-helix.	DISCUSS
+26	31	TOCA1	protein	The interhelical loops of TOCA1 and CIP4 differ from the same region in the HR1 domains of PRK1 in that they are longer and contain two short stretches of 310-helix.	DISCUSS
+36	40	CIP4	protein	The interhelical loops of TOCA1 and CIP4 differ from the same region in the HR1 domains of PRK1 in that they are longer and contain two short stretches of 310-helix.	DISCUSS
+76	79	HR1	structure_element	The interhelical loops of TOCA1 and CIP4 differ from the same region in the HR1 domains of PRK1 in that they are longer and contain two short stretches of 310-helix.	DISCUSS
+91	95	PRK1	protein	The interhelical loops of TOCA1 and CIP4 differ from the same region in the HR1 domains of PRK1 in that they are longer and contain two short stretches of 310-helix.	DISCUSS
+155	164	310-helix	structure_element	The interhelical loops of TOCA1 and CIP4 differ from the same region in the HR1 domains of PRK1 in that they are longer and contain two short stretches of 310-helix.	DISCUSS
+28	53	G protein-binding surface	site	This region lies within the G protein-binding surface of all of the HR1 domains (Fig. 4D).	DISCUSS
+68	71	HR1	structure_element	This region lies within the G protein-binding surface of all of the HR1 domains (Fig. 4D).	DISCUSS
+0	5	TOCA1	protein	TOCA1 and CIP4 both bind weakly to Cdc42, whereas the HR1a domain of PRK1 binds tightly to RhoA and Rac1, and the HR1b domain binds to Rac1.	DISCUSS
+10	14	CIP4	protein	TOCA1 and CIP4 both bind weakly to Cdc42, whereas the HR1a domain of PRK1 binds tightly to RhoA and Rac1, and the HR1b domain binds to Rac1.	DISCUSS
+35	40	Cdc42	protein	TOCA1 and CIP4 both bind weakly to Cdc42, whereas the HR1a domain of PRK1 binds tightly to RhoA and Rac1, and the HR1b domain binds to Rac1.	DISCUSS
+54	58	HR1a	structure_element	TOCA1 and CIP4 both bind weakly to Cdc42, whereas the HR1a domain of PRK1 binds tightly to RhoA and Rac1, and the HR1b domain binds to Rac1.	DISCUSS
+69	73	PRK1	protein	TOCA1 and CIP4 both bind weakly to Cdc42, whereas the HR1a domain of PRK1 binds tightly to RhoA and Rac1, and the HR1b domain binds to Rac1.	DISCUSS
+91	95	RhoA	protein	TOCA1 and CIP4 both bind weakly to Cdc42, whereas the HR1a domain of PRK1 binds tightly to RhoA and Rac1, and the HR1b domain binds to Rac1.	DISCUSS
+100	104	Rac1	protein	TOCA1 and CIP4 both bind weakly to Cdc42, whereas the HR1a domain of PRK1 binds tightly to RhoA and Rac1, and the HR1b domain binds to Rac1.	DISCUSS
+114	118	HR1b	structure_element	TOCA1 and CIP4 both bind weakly to Cdc42, whereas the HR1a domain of PRK1 binds tightly to RhoA and Rac1, and the HR1b domain binds to Rac1.	DISCUSS
+135	139	Rac1	protein	TOCA1 and CIP4 both bind weakly to Cdc42, whereas the HR1a domain of PRK1 binds tightly to RhoA and Rac1, and the HR1b domain binds to Rac1.	DISCUSS
+34	39	TOCA1	protein	The structural features shared by TOCA1 and CIP4 may therefore be related to Cdc42 binding specificity and the low affinities.	DISCUSS
+44	48	CIP4	protein	The structural features shared by TOCA1 and CIP4 may therefore be related to Cdc42 binding specificity and the low affinities.	DISCUSS
+77	82	Cdc42	protein	The structural features shared by TOCA1 and CIP4 may therefore be related to Cdc42 binding specificity and the low affinities.	DISCUSS
+3	7	free	protein_state	In free TOCA1, the side chains of the interhelical region make extensive contacts with residues in helix 1.	DISCUSS
+8	13	TOCA1	protein	In free TOCA1, the side chains of the interhelical region make extensive contacts with residues in helix 1.	DISCUSS
+38	57	interhelical region	structure_element	In free TOCA1, the side chains of the interhelical region make extensive contacts with residues in helix 1.	DISCUSS
+99	106	helix 1	structure_element	In free TOCA1, the side chains of the interhelical region make extensive contacts with residues in helix 1.	DISCUSS
+57	68	presence of	protein_state	Many of these residues are significantly affected in the presence of Cdc42, so it is likely that the conformation of this loop is altered in the Cdc42 complex.	DISCUSS
+69	74	Cdc42	protein	Many of these residues are significantly affected in the presence of Cdc42, so it is likely that the conformation of this loop is altered in the Cdc42 complex.	DISCUSS
+122	126	loop	structure_element	Many of these residues are significantly affected in the presence of Cdc42, so it is likely that the conformation of this loop is altered in the Cdc42 complex.	DISCUSS
+145	150	Cdc42	protein	Many of these residues are significantly affected in the presence of Cdc42, so it is likely that the conformation of this loop is altered in the Cdc42 complex.	DISCUSS
+67	75	mutation	experimental_method	These observations therefore provide a molecular mechanism whereby mutation of Met383-Gly384-Asp385 to Ile383-Ser384-Thr385 abolishes TOCA1 binding to Cdc42.	DISCUSS
+79	85	Met383	residue_name_number	These observations therefore provide a molecular mechanism whereby mutation of Met383-Gly384-Asp385 to Ile383-Ser384-Thr385 abolishes TOCA1 binding to Cdc42.	DISCUSS
+86	92	Gly384	residue_name_number	These observations therefore provide a molecular mechanism whereby mutation of Met383-Gly384-Asp385 to Ile383-Ser384-Thr385 abolishes TOCA1 binding to Cdc42.	DISCUSS
+93	99	Asp385	residue_name_number	These observations therefore provide a molecular mechanism whereby mutation of Met383-Gly384-Asp385 to Ile383-Ser384-Thr385 abolishes TOCA1 binding to Cdc42.	DISCUSS
+103	109	Ile383	residue_name_number	These observations therefore provide a molecular mechanism whereby mutation of Met383-Gly384-Asp385 to Ile383-Ser384-Thr385 abolishes TOCA1 binding to Cdc42.	DISCUSS
+110	116	Ser384	residue_name_number	These observations therefore provide a molecular mechanism whereby mutation of Met383-Gly384-Asp385 to Ile383-Ser384-Thr385 abolishes TOCA1 binding to Cdc42.	DISCUSS
+117	123	Thr385	residue_name_number	These observations therefore provide a molecular mechanism whereby mutation of Met383-Gly384-Asp385 to Ile383-Ser384-Thr385 abolishes TOCA1 binding to Cdc42.	DISCUSS
+134	139	TOCA1	protein	These observations therefore provide a molecular mechanism whereby mutation of Met383-Gly384-Asp385 to Ile383-Ser384-Thr385 abolishes TOCA1 binding to Cdc42.	DISCUSS
+151	156	Cdc42	protein	These observations therefore provide a molecular mechanism whereby mutation of Met383-Gly384-Asp385 to Ile383-Ser384-Thr385 abolishes TOCA1 binding to Cdc42.	DISCUSS
+18	23	model	evidence	The lowest energy model produced by HADDOCK using ambiguous interaction restraints from the titration data resembled the NMR structures of RhoA and Rac1 in complex with their HR1 domain partners.	DISCUSS
+36	43	HADDOCK	experimental_method	The lowest energy model produced by HADDOCK using ambiguous interaction restraints from the titration data resembled the NMR structures of RhoA and Rac1 in complex with their HR1 domain partners.	DISCUSS
+92	101	titration	evidence	The lowest energy model produced by HADDOCK using ambiguous interaction restraints from the titration data resembled the NMR structures of RhoA and Rac1 in complex with their HR1 domain partners.	DISCUSS
+121	124	NMR	experimental_method	The lowest energy model produced by HADDOCK using ambiguous interaction restraints from the titration data resembled the NMR structures of RhoA and Rac1 in complex with their HR1 domain partners.	DISCUSS
+125	135	structures	evidence	The lowest energy model produced by HADDOCK using ambiguous interaction restraints from the titration data resembled the NMR structures of RhoA and Rac1 in complex with their HR1 domain partners.	DISCUSS
+139	143	RhoA	protein	The lowest energy model produced by HADDOCK using ambiguous interaction restraints from the titration data resembled the NMR structures of RhoA and Rac1 in complex with their HR1 domain partners.	DISCUSS
+148	152	Rac1	protein	The lowest energy model produced by HADDOCK using ambiguous interaction restraints from the titration data resembled the NMR structures of RhoA and Rac1 in complex with their HR1 domain partners.	DISCUSS
+153	168	in complex with	protein_state	The lowest energy model produced by HADDOCK using ambiguous interaction restraints from the titration data resembled the NMR structures of RhoA and Rac1 in complex with their HR1 domain partners.	DISCUSS
+175	178	HR1	structure_element	The lowest energy model produced by HADDOCK using ambiguous interaction restraints from the titration data resembled the NMR structures of RhoA and Rac1 in complex with their HR1 domain partners.	DISCUSS
+13	19	Phe-56	residue_name_number	For example, Phe-56Cdc42, which is not visible in free Cdc42 or Cdc42·HR1TOCA1, is close to the TOCA1 HR1 (Fig. 6A).	DISCUSS
+19	24	Cdc42	protein	For example, Phe-56Cdc42, which is not visible in free Cdc42 or Cdc42·HR1TOCA1, is close to the TOCA1 HR1 (Fig. 6A).	DISCUSS
+50	54	free	protein_state	For example, Phe-56Cdc42, which is not visible in free Cdc42 or Cdc42·HR1TOCA1, is close to the TOCA1 HR1 (Fig. 6A).	DISCUSS
+55	60	Cdc42	protein	For example, Phe-56Cdc42, which is not visible in free Cdc42 or Cdc42·HR1TOCA1, is close to the TOCA1 HR1 (Fig. 6A).	DISCUSS
+64	78	Cdc42·HR1TOCA1	complex_assembly	For example, Phe-56Cdc42, which is not visible in free Cdc42 or Cdc42·HR1TOCA1, is close to the TOCA1 HR1 (Fig. 6A).	DISCUSS
+96	101	TOCA1	protein	For example, Phe-56Cdc42, which is not visible in free Cdc42 or Cdc42·HR1TOCA1, is close to the TOCA1 HR1 (Fig. 6A).	DISCUSS
+102	105	HR1	structure_element	For example, Phe-56Cdc42, which is not visible in free Cdc42 or Cdc42·HR1TOCA1, is close to the TOCA1 HR1 (Fig. 6A).	DISCUSS
+0	6	Phe-56	residue_name_number	Phe-56Cdc42, which is a Trp in both Rac1 and RhoA (Fig. 6C), is thought to pack behind switch I when Cdc42 interacts with ACK, maintaining the switch in a binding-competent orientation.	DISCUSS
+6	11	Cdc42	protein	Phe-56Cdc42, which is a Trp in both Rac1 and RhoA (Fig. 6C), is thought to pack behind switch I when Cdc42 interacts with ACK, maintaining the switch in a binding-competent orientation.	DISCUSS
+24	27	Trp	residue_name	Phe-56Cdc42, which is a Trp in both Rac1 and RhoA (Fig. 6C), is thought to pack behind switch I when Cdc42 interacts with ACK, maintaining the switch in a binding-competent orientation.	DISCUSS
+36	40	Rac1	protein	Phe-56Cdc42, which is a Trp in both Rac1 and RhoA (Fig. 6C), is thought to pack behind switch I when Cdc42 interacts with ACK, maintaining the switch in a binding-competent orientation.	DISCUSS
+45	49	RhoA	protein	Phe-56Cdc42, which is a Trp in both Rac1 and RhoA (Fig. 6C), is thought to pack behind switch I when Cdc42 interacts with ACK, maintaining the switch in a binding-competent orientation.	DISCUSS
+87	95	switch I	site	Phe-56Cdc42, which is a Trp in both Rac1 and RhoA (Fig. 6C), is thought to pack behind switch I when Cdc42 interacts with ACK, maintaining the switch in a binding-competent orientation.	DISCUSS
+101	106	Cdc42	protein	Phe-56Cdc42, which is a Trp in both Rac1 and RhoA (Fig. 6C), is thought to pack behind switch I when Cdc42 interacts with ACK, maintaining the switch in a binding-competent orientation.	DISCUSS
+122	125	ACK	protein	Phe-56Cdc42, which is a Trp in both Rac1 and RhoA (Fig. 6C), is thought to pack behind switch I when Cdc42 interacts with ACK, maintaining the switch in a binding-competent orientation.	DISCUSS
+55	60	Cdc42	protein	This residue has also been identified as important for Cdc42-WASP binding.	DISCUSS
+61	65	WASP	protein	This residue has also been identified as important for Cdc42-WASP binding.	DISCUSS
+0	6	Phe-56	residue_name_number	Phe-56Cdc42 is therefore likely to be involved in the Cdc42-TOCA1 interaction, probably by stabilizing the position of switch I.	DISCUSS
+6	11	Cdc42	protein	Phe-56Cdc42 is therefore likely to be involved in the Cdc42-TOCA1 interaction, probably by stabilizing the position of switch I.	DISCUSS
+54	59	Cdc42	protein	Phe-56Cdc42 is therefore likely to be involved in the Cdc42-TOCA1 interaction, probably by stabilizing the position of switch I.	DISCUSS
+60	65	TOCA1	protein	Phe-56Cdc42 is therefore likely to be involved in the Cdc42-TOCA1 interaction, probably by stabilizing the position of switch I.	DISCUSS
+119	127	switch I	site	Phe-56Cdc42 is therefore likely to be involved in the Cdc42-TOCA1 interaction, probably by stabilizing the position of switch I.	DISCUSS
+39	53	Cdc42·HR1TOCA1	complex_assembly	Some residues that are affected in the Cdc42·HR1TOCA1 complex but do not correspond to contact residues of RhoA or Rac1 (Fig. 6C) may contact HR1TOCA1 directly (Fig. 6D).	DISCUSS
+107	111	RhoA	protein	Some residues that are affected in the Cdc42·HR1TOCA1 complex but do not correspond to contact residues of RhoA or Rac1 (Fig. 6C) may contact HR1TOCA1 directly (Fig. 6D).	DISCUSS
+115	119	Rac1	protein	Some residues that are affected in the Cdc42·HR1TOCA1 complex but do not correspond to contact residues of RhoA or Rac1 (Fig. 6C) may contact HR1TOCA1 directly (Fig. 6D).	DISCUSS
+142	145	HR1	structure_element	Some residues that are affected in the Cdc42·HR1TOCA1 complex but do not correspond to contact residues of RhoA or Rac1 (Fig. 6C) may contact HR1TOCA1 directly (Fig. 6D).	DISCUSS
+145	150	TOCA1	protein	Some residues that are affected in the Cdc42·HR1TOCA1 complex but do not correspond to contact residues of RhoA or Rac1 (Fig. 6C) may contact HR1TOCA1 directly (Fig. 6D).	DISCUSS
+0	5	Gln-2	residue_name_number	Gln-2Cdc42, which has also been identified as a contact residue in the Cdc42·ACK complex, contacts Val-376TOCA1 and Asn-380TOCA1 in the model and disrupts the contacts between the interhelical loop and the first helix of the TOCA1 coiled-coil.	DISCUSS
+5	10	Cdc42	protein	Gln-2Cdc42, which has also been identified as a contact residue in the Cdc42·ACK complex, contacts Val-376TOCA1 and Asn-380TOCA1 in the model and disrupts the contacts between the interhelical loop and the first helix of the TOCA1 coiled-coil.	DISCUSS
+71	80	Cdc42·ACK	complex_assembly	Gln-2Cdc42, which has also been identified as a contact residue in the Cdc42·ACK complex, contacts Val-376TOCA1 and Asn-380TOCA1 in the model and disrupts the contacts between the interhelical loop and the first helix of the TOCA1 coiled-coil.	DISCUSS
+99	106	Val-376	residue_name_number	Gln-2Cdc42, which has also been identified as a contact residue in the Cdc42·ACK complex, contacts Val-376TOCA1 and Asn-380TOCA1 in the model and disrupts the contacts between the interhelical loop and the first helix of the TOCA1 coiled-coil.	DISCUSS
+106	111	TOCA1	protein	Gln-2Cdc42, which has also been identified as a contact residue in the Cdc42·ACK complex, contacts Val-376TOCA1 and Asn-380TOCA1 in the model and disrupts the contacts between the interhelical loop and the first helix of the TOCA1 coiled-coil.	DISCUSS
+116	123	Asn-380	residue_name_number	Gln-2Cdc42, which has also been identified as a contact residue in the Cdc42·ACK complex, contacts Val-376TOCA1 and Asn-380TOCA1 in the model and disrupts the contacts between the interhelical loop and the first helix of the TOCA1 coiled-coil.	DISCUSS
+123	128	TOCA1	protein	Gln-2Cdc42, which has also been identified as a contact residue in the Cdc42·ACK complex, contacts Val-376TOCA1 and Asn-380TOCA1 in the model and disrupts the contacts between the interhelical loop and the first helix of the TOCA1 coiled-coil.	DISCUSS
+180	197	interhelical loop	structure_element	Gln-2Cdc42, which has also been identified as a contact residue in the Cdc42·ACK complex, contacts Val-376TOCA1 and Asn-380TOCA1 in the model and disrupts the contacts between the interhelical loop and the first helix of the TOCA1 coiled-coil.	DISCUSS
+206	217	first helix	structure_element	Gln-2Cdc42, which has also been identified as a contact residue in the Cdc42·ACK complex, contacts Val-376TOCA1 and Asn-380TOCA1 in the model and disrupts the contacts between the interhelical loop and the first helix of the TOCA1 coiled-coil.	DISCUSS
+225	230	TOCA1	protein	Gln-2Cdc42, which has also been identified as a contact residue in the Cdc42·ACK complex, contacts Val-376TOCA1 and Asn-380TOCA1 in the model and disrupts the contacts between the interhelical loop and the first helix of the TOCA1 coiled-coil.	DISCUSS
+231	242	coiled-coil	structure_element	Gln-2Cdc42, which has also been identified as a contact residue in the Cdc42·ACK complex, contacts Val-376TOCA1 and Asn-380TOCA1 in the model and disrupts the contacts between the interhelical loop and the first helix of the TOCA1 coiled-coil.	DISCUSS
+0	6	Thr-52	residue_name_number	Thr-52Cdc42, which has also been identified as making minor contacts with ACK, falls near the side chains of HR1TOCA1 helix 1, particularly Lys-372TOCA1, whereas the equivalent position in Rac1 is Asn-52Rac1.	DISCUSS
+6	11	Cdc42	protein	Thr-52Cdc42, which has also been identified as making minor contacts with ACK, falls near the side chains of HR1TOCA1 helix 1, particularly Lys-372TOCA1, whereas the equivalent position in Rac1 is Asn-52Rac1.	DISCUSS
+74	77	ACK	protein	Thr-52Cdc42, which has also been identified as making minor contacts with ACK, falls near the side chains of HR1TOCA1 helix 1, particularly Lys-372TOCA1, whereas the equivalent position in Rac1 is Asn-52Rac1.	DISCUSS
+109	112	HR1	structure_element	Thr-52Cdc42, which has also been identified as making minor contacts with ACK, falls near the side chains of HR1TOCA1 helix 1, particularly Lys-372TOCA1, whereas the equivalent position in Rac1 is Asn-52Rac1.	DISCUSS
+112	117	TOCA1	protein	Thr-52Cdc42, which has also been identified as making minor contacts with ACK, falls near the side chains of HR1TOCA1 helix 1, particularly Lys-372TOCA1, whereas the equivalent position in Rac1 is Asn-52Rac1.	DISCUSS
+118	125	helix 1	structure_element	Thr-52Cdc42, which has also been identified as making minor contacts with ACK, falls near the side chains of HR1TOCA1 helix 1, particularly Lys-372TOCA1, whereas the equivalent position in Rac1 is Asn-52Rac1.	DISCUSS
+140	147	Lys-372	residue_name_number	Thr-52Cdc42, which has also been identified as making minor contacts with ACK, falls near the side chains of HR1TOCA1 helix 1, particularly Lys-372TOCA1, whereas the equivalent position in Rac1 is Asn-52Rac1.	DISCUSS
+147	152	TOCA1	protein	Thr-52Cdc42, which has also been identified as making minor contacts with ACK, falls near the side chains of HR1TOCA1 helix 1, particularly Lys-372TOCA1, whereas the equivalent position in Rac1 is Asn-52Rac1.	DISCUSS
+189	193	Rac1	protein	Thr-52Cdc42, which has also been identified as making minor contacts with ACK, falls near the side chains of HR1TOCA1 helix 1, particularly Lys-372TOCA1, whereas the equivalent position in Rac1 is Asn-52Rac1.	DISCUSS
+197	203	Asn-52	residue_name_number	Thr-52Cdc42, which has also been identified as making minor contacts with ACK, falls near the side chains of HR1TOCA1 helix 1, particularly Lys-372TOCA1, whereas the equivalent position in Rac1 is Asn-52Rac1.	DISCUSS
+203	207	Rac1	protein	Thr-52Cdc42, which has also been identified as making minor contacts with ACK, falls near the side chains of HR1TOCA1 helix 1, particularly Lys-372TOCA1, whereas the equivalent position in Rac1 is Asn-52Rac1.	DISCUSS
+0	4	N52T	mutant	N52T is one of a combination of seven residues found to confer ACK binding on Rac1 and so may represent a specific Cdc42-effector contact residue.	DISCUSS
+63	66	ACK	protein	N52T is one of a combination of seven residues found to confer ACK binding on Rac1 and so may represent a specific Cdc42-effector contact residue.	DISCUSS
+78	82	Rac1	protein	N52T is one of a combination of seven residues found to confer ACK binding on Rac1 and so may represent a specific Cdc42-effector contact residue.	DISCUSS
+115	120	Cdc42	protein	N52T is one of a combination of seven residues found to confer ACK binding on Rac1 and so may represent a specific Cdc42-effector contact residue.	DISCUSS
+27	34	Lys-372	residue_name_number	The position equivalent to Lys-372TOCA1 in PRK1 is Glu-58HR1a or Gln-151HR1b.	DISCUSS
+34	39	TOCA1	protein	The position equivalent to Lys-372TOCA1 in PRK1 is Glu-58HR1a or Gln-151HR1b.	DISCUSS
+43	47	PRK1	protein	The position equivalent to Lys-372TOCA1 in PRK1 is Glu-58HR1a or Gln-151HR1b.	DISCUSS
+51	57	Glu-58	residue_name_number	The position equivalent to Lys-372TOCA1 in PRK1 is Glu-58HR1a or Gln-151HR1b.	DISCUSS
+57	61	HR1a	structure_element	The position equivalent to Lys-372TOCA1 in PRK1 is Glu-58HR1a or Gln-151HR1b.	DISCUSS
+65	72	Gln-151	residue_name_number	The position equivalent to Lys-372TOCA1 in PRK1 is Glu-58HR1a or Gln-151HR1b.	DISCUSS
+72	76	HR1b	structure_element	The position equivalent to Lys-372TOCA1 in PRK1 is Glu-58HR1a or Gln-151HR1b.	DISCUSS
+0	6	Thr-52	residue_name_number	Thr-52Cdc42-Lys-372TOCA1 may therefore represent a specific Cdc42-HR1TOCA1 contact.	DISCUSS
+6	11	Cdc42	protein	Thr-52Cdc42-Lys-372TOCA1 may therefore represent a specific Cdc42-HR1TOCA1 contact.	DISCUSS
+12	19	Lys-372	residue_name_number	Thr-52Cdc42-Lys-372TOCA1 may therefore represent a specific Cdc42-HR1TOCA1 contact.	DISCUSS
+19	24	TOCA1	protein	Thr-52Cdc42-Lys-372TOCA1 may therefore represent a specific Cdc42-HR1TOCA1 contact.	DISCUSS
+60	65	Cdc42	protein	Thr-52Cdc42-Lys-372TOCA1 may therefore represent a specific Cdc42-HR1TOCA1 contact.	DISCUSS
+66	69	HR1	structure_element	Thr-52Cdc42-Lys-372TOCA1 may therefore represent a specific Cdc42-HR1TOCA1 contact.	DISCUSS
+69	74	TOCA1	protein	Thr-52Cdc42-Lys-372TOCA1 may therefore represent a specific Cdc42-HR1TOCA1 contact.	DISCUSS
+0	6	Arg-68	residue_name_number	Arg-68Cdc42 of switch II is positioned close to Glu-395TOCA1 (Fig. 6D), suggesting a direct electrostatic contact between switch II of Cdc42 and helix 2 of the HR1 domain.	DISCUSS
+6	11	Cdc42	protein	Arg-68Cdc42 of switch II is positioned close to Glu-395TOCA1 (Fig. 6D), suggesting a direct electrostatic contact between switch II of Cdc42 and helix 2 of the HR1 domain.	DISCUSS
+15	24	switch II	site	Arg-68Cdc42 of switch II is positioned close to Glu-395TOCA1 (Fig. 6D), suggesting a direct electrostatic contact between switch II of Cdc42 and helix 2 of the HR1 domain.	DISCUSS
+48	55	Glu-395	residue_name_number	Arg-68Cdc42 of switch II is positioned close to Glu-395TOCA1 (Fig. 6D), suggesting a direct electrostatic contact between switch II of Cdc42 and helix 2 of the HR1 domain.	DISCUSS
+55	60	TOCA1	protein	Arg-68Cdc42 of switch II is positioned close to Glu-395TOCA1 (Fig. 6D), suggesting a direct electrostatic contact between switch II of Cdc42 and helix 2 of the HR1 domain.	DISCUSS
+122	131	switch II	site	Arg-68Cdc42 of switch II is positioned close to Glu-395TOCA1 (Fig. 6D), suggesting a direct electrostatic contact between switch II of Cdc42 and helix 2 of the HR1 domain.	DISCUSS
+135	140	Cdc42	protein	Arg-68Cdc42 of switch II is positioned close to Glu-395TOCA1 (Fig. 6D), suggesting a direct electrostatic contact between switch II of Cdc42 and helix 2 of the HR1 domain.	DISCUSS
+145	152	helix 2	structure_element	Arg-68Cdc42 of switch II is positioned close to Glu-395TOCA1 (Fig. 6D), suggesting a direct electrostatic contact between switch II of Cdc42 and helix 2 of the HR1 domain.	DISCUSS
+160	163	HR1	structure_element	Arg-68Cdc42 of switch II is positioned close to Glu-395TOCA1 (Fig. 6D), suggesting a direct electrostatic contact between switch II of Cdc42 and helix 2 of the HR1 domain.	DISCUSS
+15	18	Arg	residue_name	The equivalent Arg in Rac1 and RhoA is pointing away from the HR1 domains of PRK1.	DISCUSS
+22	26	Rac1	protein	The equivalent Arg in Rac1 and RhoA is pointing away from the HR1 domains of PRK1.	DISCUSS
+31	35	RhoA	protein	The equivalent Arg in Rac1 and RhoA is pointing away from the HR1 domains of PRK1.	DISCUSS
+62	65	HR1	structure_element	The equivalent Arg in Rac1 and RhoA is pointing away from the HR1 domains of PRK1.	DISCUSS
+77	81	PRK1	protein	The equivalent Arg in Rac1 and RhoA is pointing away from the HR1 domains of PRK1.	DISCUSS
+38	43	Cdc42	protein	The importance of this residue in the Cdc42-TOCA1 interaction remains unclear, although its mutation reduces binding to RhoGAP, suggesting that it can be involved in Cdc42 interactions.	DISCUSS
+44	49	TOCA1	protein	The importance of this residue in the Cdc42-TOCA1 interaction remains unclear, although its mutation reduces binding to RhoGAP, suggesting that it can be involved in Cdc42 interactions.	DISCUSS
+92	100	mutation	experimental_method	The importance of this residue in the Cdc42-TOCA1 interaction remains unclear, although its mutation reduces binding to RhoGAP, suggesting that it can be involved in Cdc42 interactions.	DISCUSS
+120	126	RhoGAP	protein	The importance of this residue in the Cdc42-TOCA1 interaction remains unclear, although its mutation reduces binding to RhoGAP, suggesting that it can be involved in Cdc42 interactions.	DISCUSS
+166	171	Cdc42	protein	The importance of this residue in the Cdc42-TOCA1 interaction remains unclear, although its mutation reduces binding to RhoGAP, suggesting that it can be involved in Cdc42 interactions.	DISCUSS
+4	22	solution structure	evidence	The solution structure of the TOCA1 HR1 domain presented here, along with the model of the HR1TOCA1·Cdc42 complex is consistent with a conserved mode of binding across the known HR1 domain-Rho family interactions, despite their differing affinities.	DISCUSS
+30	35	TOCA1	protein	The solution structure of the TOCA1 HR1 domain presented here, along with the model of the HR1TOCA1·Cdc42 complex is consistent with a conserved mode of binding across the known HR1 domain-Rho family interactions, despite their differing affinities.	DISCUSS
+36	39	HR1	structure_element	The solution structure of the TOCA1 HR1 domain presented here, along with the model of the HR1TOCA1·Cdc42 complex is consistent with a conserved mode of binding across the known HR1 domain-Rho family interactions, despite their differing affinities.	DISCUSS
+91	105	HR1TOCA1·Cdc42	complex_assembly	The solution structure of the TOCA1 HR1 domain presented here, along with the model of the HR1TOCA1·Cdc42 complex is consistent with a conserved mode of binding across the known HR1 domain-Rho family interactions, despite their differing affinities.	DISCUSS
+178	181	HR1	structure_element	The solution structure of the TOCA1 HR1 domain presented here, along with the model of the HR1TOCA1·Cdc42 complex is consistent with a conserved mode of binding across the known HR1 domain-Rho family interactions, despite their differing affinities.	DISCUSS
+36	72	structural and thermodynamic studies	experimental_method	The weak binding prevented detailed structural and thermodynamic studies of the complex.	DISCUSS
+13	31	structural studies	experimental_method	Nonetheless, structural studies of the TOCA1 HR1 domain, combined with chemical shift mapping, have highlighted some potentially interesting differences between Cdc42-HR1TOCA1 and RhoA/Rac1-HR1PRK1 binding.	DISCUSS
+39	44	TOCA1	protein	Nonetheless, structural studies of the TOCA1 HR1 domain, combined with chemical shift mapping, have highlighted some potentially interesting differences between Cdc42-HR1TOCA1 and RhoA/Rac1-HR1PRK1 binding.	DISCUSS
+45	48	HR1	structure_element	Nonetheless, structural studies of the TOCA1 HR1 domain, combined with chemical shift mapping, have highlighted some potentially interesting differences between Cdc42-HR1TOCA1 and RhoA/Rac1-HR1PRK1 binding.	DISCUSS
+71	93	chemical shift mapping	experimental_method	Nonetheless, structural studies of the TOCA1 HR1 domain, combined with chemical shift mapping, have highlighted some potentially interesting differences between Cdc42-HR1TOCA1 and RhoA/Rac1-HR1PRK1 binding.	DISCUSS
+161	166	Cdc42	protein	Nonetheless, structural studies of the TOCA1 HR1 domain, combined with chemical shift mapping, have highlighted some potentially interesting differences between Cdc42-HR1TOCA1 and RhoA/Rac1-HR1PRK1 binding.	DISCUSS
+167	170	HR1	structure_element	Nonetheless, structural studies of the TOCA1 HR1 domain, combined with chemical shift mapping, have highlighted some potentially interesting differences between Cdc42-HR1TOCA1 and RhoA/Rac1-HR1PRK1 binding.	DISCUSS
+170	175	TOCA1	protein	Nonetheless, structural studies of the TOCA1 HR1 domain, combined with chemical shift mapping, have highlighted some potentially interesting differences between Cdc42-HR1TOCA1 and RhoA/Rac1-HR1PRK1 binding.	DISCUSS
+180	184	RhoA	protein	Nonetheless, structural studies of the TOCA1 HR1 domain, combined with chemical shift mapping, have highlighted some potentially interesting differences between Cdc42-HR1TOCA1 and RhoA/Rac1-HR1PRK1 binding.	DISCUSS
+185	189	Rac1	protein	Nonetheless, structural studies of the TOCA1 HR1 domain, combined with chemical shift mapping, have highlighted some potentially interesting differences between Cdc42-HR1TOCA1 and RhoA/Rac1-HR1PRK1 binding.	DISCUSS
+190	193	HR1	structure_element	Nonetheless, structural studies of the TOCA1 HR1 domain, combined with chemical shift mapping, have highlighted some potentially interesting differences between Cdc42-HR1TOCA1 and RhoA/Rac1-HR1PRK1 binding.	DISCUSS
+193	197	PRK1	protein	Nonetheless, structural studies of the TOCA1 HR1 domain, combined with chemical shift mapping, have highlighted some potentially interesting differences between Cdc42-HR1TOCA1 and RhoA/Rac1-HR1PRK1 binding.	DISCUSS
+63	66	HR1	structure_element	We have previously postulated that the inherent flexibility of HR1 domains contributes to their ability to bind to different Rho family G proteins, with Rho-binding HR1 domains displaying increased flexibility, reflected in their lower melting temperatures (Tm) and Rac binders being more rigid.	DISCUSS
+125	146	Rho family G proteins	protein_type	We have previously postulated that the inherent flexibility of HR1 domains contributes to their ability to bind to different Rho family G proteins, with Rho-binding HR1 domains displaying increased flexibility, reflected in their lower melting temperatures (Tm) and Rac binders being more rigid.	DISCUSS
+165	168	HR1	structure_element	We have previously postulated that the inherent flexibility of HR1 domains contributes to their ability to bind to different Rho family G proteins, with Rho-binding HR1 domains displaying increased flexibility, reflected in their lower melting temperatures (Tm) and Rac binders being more rigid.	DISCUSS
+236	256	melting temperatures	evidence	We have previously postulated that the inherent flexibility of HR1 domains contributes to their ability to bind to different Rho family G proteins, with Rho-binding HR1 domains displaying increased flexibility, reflected in their lower melting temperatures (Tm) and Rac binders being more rigid.	DISCUSS
+258	260	Tm	evidence	We have previously postulated that the inherent flexibility of HR1 domains contributes to their ability to bind to different Rho family G proteins, with Rho-binding HR1 domains displaying increased flexibility, reflected in their lower melting temperatures (Tm) and Rac binders being more rigid.	DISCUSS
+266	269	Rac	protein_type	We have previously postulated that the inherent flexibility of HR1 domains contributes to their ability to bind to different Rho family G proteins, with Rho-binding HR1 domains displaying increased flexibility, reflected in their lower melting temperatures (Tm) and Rac binders being more rigid.	DISCUSS
+4	6	Tm	evidence	The Tm of the TOCA1 HR1 domain is 61.9 °C (data not shown), which is the highest Tm that we have measured for an HR1 domain thus far.	DISCUSS
+14	19	TOCA1	protein	The Tm of the TOCA1 HR1 domain is 61.9 °C (data not shown), which is the highest Tm that we have measured for an HR1 domain thus far.	DISCUSS
+20	23	HR1	structure_element	The Tm of the TOCA1 HR1 domain is 61.9 °C (data not shown), which is the highest Tm that we have measured for an HR1 domain thus far.	DISCUSS
+81	83	Tm	evidence	The Tm of the TOCA1 HR1 domain is 61.9 °C (data not shown), which is the highest Tm that we have measured for an HR1 domain thus far.	DISCUSS
+113	116	HR1	structure_element	The Tm of the TOCA1 HR1 domain is 61.9 °C (data not shown), which is the highest Tm that we have measured for an HR1 domain thus far.	DISCUSS
+28	33	TOCA1	protein	As such, the ability of the TOCA1 HR1 domain to bind to Cdc42 (a close relative of Rac1 rather than RhoA) fits this trend.	DISCUSS
+34	37	HR1	structure_element	As such, the ability of the TOCA1 HR1 domain to bind to Cdc42 (a close relative of Rac1 rather than RhoA) fits this trend.	DISCUSS
+56	61	Cdc42	protein	As such, the ability of the TOCA1 HR1 domain to bind to Cdc42 (a close relative of Rac1 rather than RhoA) fits this trend.	DISCUSS
+83	87	Rac1	protein	As such, the ability of the TOCA1 HR1 domain to bind to Cdc42 (a close relative of Rac1 rather than RhoA) fits this trend.	DISCUSS
+100	104	RhoA	protein	As such, the ability of the TOCA1 HR1 domain to bind to Cdc42 (a close relative of Rac1 rather than RhoA) fits this trend.	DISCUSS
+61	86	G protein-binding regions	site	An investigation into the local motions, particularly in the G protein-binding regions, may offer further insight into the differential specificities and affinities of G protein-HR1 domain interactions.	DISCUSS
+168	177	G protein	protein_type	An investigation into the local motions, particularly in the G protein-binding regions, may offer further insight into the differential specificities and affinities of G protein-HR1 domain interactions.	DISCUSS
+178	181	HR1	structure_element	An investigation into the local motions, particularly in the G protein-binding regions, may offer further insight into the differential specificities and affinities of G protein-HR1 domain interactions.	DISCUSS
+24	29	Cdc42	protein	The low affinity of the Cdc42-HR1TOCA1 interaction is consistent with a tightly spatially and temporally regulated pathway, requiring combinatorial signals leading to a series of coincident weak interactions that elicit full activation.	DISCUSS
+30	33	HR1	structure_element	The low affinity of the Cdc42-HR1TOCA1 interaction is consistent with a tightly spatially and temporally regulated pathway, requiring combinatorial signals leading to a series of coincident weak interactions that elicit full activation.	DISCUSS
+33	38	TOCA1	protein	The low affinity of the Cdc42-HR1TOCA1 interaction is consistent with a tightly spatially and temporally regulated pathway, requiring combinatorial signals leading to a series of coincident weak interactions that elicit full activation.	DISCUSS
+4	7	HR1	structure_element	The HR1 domains from other TOCA family members, CIP4 and FBP17, also bind at low micromolar affinities to Cdc42, so the low affinity interaction appears to be commonplace among this family of HR1 domain proteins, in contrast to the PRK family.	DISCUSS
+27	46	TOCA family members	protein_type	The HR1 domains from other TOCA family members, CIP4 and FBP17, also bind at low micromolar affinities to Cdc42, so the low affinity interaction appears to be commonplace among this family of HR1 domain proteins, in contrast to the PRK family.	DISCUSS
+48	52	CIP4	protein	The HR1 domains from other TOCA family members, CIP4 and FBP17, also bind at low micromolar affinities to Cdc42, so the low affinity interaction appears to be commonplace among this family of HR1 domain proteins, in contrast to the PRK family.	DISCUSS
+57	62	FBP17	protein	The HR1 domains from other TOCA family members, CIP4 and FBP17, also bind at low micromolar affinities to Cdc42, so the low affinity interaction appears to be commonplace among this family of HR1 domain proteins, in contrast to the PRK family.	DISCUSS
+106	111	Cdc42	protein	The HR1 domains from other TOCA family members, CIP4 and FBP17, also bind at low micromolar affinities to Cdc42, so the low affinity interaction appears to be commonplace among this family of HR1 domain proteins, in contrast to the PRK family.	DISCUSS
+192	211	HR1 domain proteins	protein_type	The HR1 domains from other TOCA family members, CIP4 and FBP17, also bind at low micromolar affinities to Cdc42, so the low affinity interaction appears to be commonplace among this family of HR1 domain proteins, in contrast to the PRK family.	DISCUSS
+232	242	PRK family	protein_type	The HR1 domains from other TOCA family members, CIP4 and FBP17, also bind at low micromolar affinities to Cdc42, so the low affinity interaction appears to be commonplace among this family of HR1 domain proteins, in contrast to the PRK family.	DISCUSS
+24	32	HR1TOCA1	structure_element	The low affinity of the HR1TOCA1-Cdc42 interaction in the context of the physiological concentration of TOCA1 in Xenopus extracts (∼10 nm) suggests that binding between TOCA1 and Cdc42 is likely to occur in vivo only when TOCA1 is at high local concentrations and membrane-localized and therefore in close proximity to activated Cdc42.	DISCUSS
+33	38	Cdc42	protein	The low affinity of the HR1TOCA1-Cdc42 interaction in the context of the physiological concentration of TOCA1 in Xenopus extracts (∼10 nm) suggests that binding between TOCA1 and Cdc42 is likely to occur in vivo only when TOCA1 is at high local concentrations and membrane-localized and therefore in close proximity to activated Cdc42.	DISCUSS
+104	109	TOCA1	protein	The low affinity of the HR1TOCA1-Cdc42 interaction in the context of the physiological concentration of TOCA1 in Xenopus extracts (∼10 nm) suggests that binding between TOCA1 and Cdc42 is likely to occur in vivo only when TOCA1 is at high local concentrations and membrane-localized and therefore in close proximity to activated Cdc42.	DISCUSS
+113	120	Xenopus	taxonomy_domain	The low affinity of the HR1TOCA1-Cdc42 interaction in the context of the physiological concentration of TOCA1 in Xenopus extracts (∼10 nm) suggests that binding between TOCA1 and Cdc42 is likely to occur in vivo only when TOCA1 is at high local concentrations and membrane-localized and therefore in close proximity to activated Cdc42.	DISCUSS
+169	174	TOCA1	protein	The low affinity of the HR1TOCA1-Cdc42 interaction in the context of the physiological concentration of TOCA1 in Xenopus extracts (∼10 nm) suggests that binding between TOCA1 and Cdc42 is likely to occur in vivo only when TOCA1 is at high local concentrations and membrane-localized and therefore in close proximity to activated Cdc42.	DISCUSS
+179	184	Cdc42	protein	The low affinity of the HR1TOCA1-Cdc42 interaction in the context of the physiological concentration of TOCA1 in Xenopus extracts (∼10 nm) suggests that binding between TOCA1 and Cdc42 is likely to occur in vivo only when TOCA1 is at high local concentrations and membrane-localized and therefore in close proximity to activated Cdc42.	DISCUSS
+222	227	TOCA1	protein	The low affinity of the HR1TOCA1-Cdc42 interaction in the context of the physiological concentration of TOCA1 in Xenopus extracts (∼10 nm) suggests that binding between TOCA1 and Cdc42 is likely to occur in vivo only when TOCA1 is at high local concentrations and membrane-localized and therefore in close proximity to activated Cdc42.	DISCUSS
+319	328	activated	protein_state	The low affinity of the HR1TOCA1-Cdc42 interaction in the context of the physiological concentration of TOCA1 in Xenopus extracts (∼10 nm) suggests that binding between TOCA1 and Cdc42 is likely to occur in vivo only when TOCA1 is at high local concentrations and membrane-localized and therefore in close proximity to activated Cdc42.	DISCUSS
+329	334	Cdc42	protein	The low affinity of the HR1TOCA1-Cdc42 interaction in the context of the physiological concentration of TOCA1 in Xenopus extracts (∼10 nm) suggests that binding between TOCA1 and Cdc42 is likely to occur in vivo only when TOCA1 is at high local concentrations and membrane-localized and therefore in close proximity to activated Cdc42.	DISCUSS
+27	38	TOCA family	protein_type	Evidence suggests that the TOCA family of proteins are recruited to the membrane via an interaction between their F-BAR domain and specific signaling lipids.	DISCUSS
+114	119	F-BAR	structure_element	Evidence suggests that the TOCA family of proteins are recruited to the membrane via an interaction between their F-BAR domain and specific signaling lipids.	DISCUSS
+52	57	F-BAR	structure_element	For example, electrostatic interactions between the F-BAR domain and the membrane are required for TOCA1 recruitment to membrane vesicles and tubules, and TOCA1-dependent actin polymerization is known to depend specifically on PI(4,5)P2.	DISCUSS
+99	104	TOCA1	protein	For example, electrostatic interactions between the F-BAR domain and the membrane are required for TOCA1 recruitment to membrane vesicles and tubules, and TOCA1-dependent actin polymerization is known to depend specifically on PI(4,5)P2.	DISCUSS
+155	160	TOCA1	protein	For example, electrostatic interactions between the F-BAR domain and the membrane are required for TOCA1 recruitment to membrane vesicles and tubules, and TOCA1-dependent actin polymerization is known to depend specifically on PI(4,5)P2.	DISCUSS
+227	236	PI(4,5)P2	chemical	For example, electrostatic interactions between the F-BAR domain and the membrane are required for TOCA1 recruitment to membrane vesicles and tubules, and TOCA1-dependent actin polymerization is known to depend specifically on PI(4,5)P2.	DISCUSS
+17	25	isolated	experimental_method	Furthermore, the isolated F-BAR domain of FBP17 has been shown to induce membrane tubulation of brain liposomes and BAR domain proteins that promote tubulation cluster on membranes at high densities.	DISCUSS
+26	31	F-BAR	structure_element	Furthermore, the isolated F-BAR domain of FBP17 has been shown to induce membrane tubulation of brain liposomes and BAR domain proteins that promote tubulation cluster on membranes at high densities.	DISCUSS
+42	47	FBP17	protein	Furthermore, the isolated F-BAR domain of FBP17 has been shown to induce membrane tubulation of brain liposomes and BAR domain proteins that promote tubulation cluster on membranes at high densities.	DISCUSS
+116	119	BAR	structure_element	Furthermore, the isolated F-BAR domain of FBP17 has been shown to induce membrane tubulation of brain liposomes and BAR domain proteins that promote tubulation cluster on membranes at high densities.	DISCUSS
+51	56	TOCA1	protein	Once at the membrane, high local concentrations of TOCA1 could exceed the Kd of F-BAR dimerization (likely to be comparable with that of the FCHo2 F-BAR domain (2.5 μm)) and that of the Cdc42-HR1TOCA1 interaction.	DISCUSS
+74	76	Kd	evidence	Once at the membrane, high local concentrations of TOCA1 could exceed the Kd of F-BAR dimerization (likely to be comparable with that of the FCHo2 F-BAR domain (2.5 μm)) and that of the Cdc42-HR1TOCA1 interaction.	DISCUSS
+80	85	F-BAR	structure_element	Once at the membrane, high local concentrations of TOCA1 could exceed the Kd of F-BAR dimerization (likely to be comparable with that of the FCHo2 F-BAR domain (2.5 μm)) and that of the Cdc42-HR1TOCA1 interaction.	DISCUSS
+86	91	dimer	oligomeric_state	Once at the membrane, high local concentrations of TOCA1 could exceed the Kd of F-BAR dimerization (likely to be comparable with that of the FCHo2 F-BAR domain (2.5 μm)) and that of the Cdc42-HR1TOCA1 interaction.	DISCUSS
+141	146	FCHo2	protein	Once at the membrane, high local concentrations of TOCA1 could exceed the Kd of F-BAR dimerization (likely to be comparable with that of the FCHo2 F-BAR domain (2.5 μm)) and that of the Cdc42-HR1TOCA1 interaction.	DISCUSS
+147	152	F-BAR	structure_element	Once at the membrane, high local concentrations of TOCA1 could exceed the Kd of F-BAR dimerization (likely to be comparable with that of the FCHo2 F-BAR domain (2.5 μm)) and that of the Cdc42-HR1TOCA1 interaction.	DISCUSS
+186	191	Cdc42	protein	Once at the membrane, high local concentrations of TOCA1 could exceed the Kd of F-BAR dimerization (likely to be comparable with that of the FCHo2 F-BAR domain (2.5 μm)) and that of the Cdc42-HR1TOCA1 interaction.	DISCUSS
+192	195	HR1	structure_element	Once at the membrane, high local concentrations of TOCA1 could exceed the Kd of F-BAR dimerization (likely to be comparable with that of the FCHo2 F-BAR domain (2.5 μm)) and that of the Cdc42-HR1TOCA1 interaction.	DISCUSS
+195	200	TOCA1	protein	Once at the membrane, high local concentrations of TOCA1 could exceed the Kd of F-BAR dimerization (likely to be comparable with that of the FCHo2 F-BAR domain (2.5 μm)) and that of the Cdc42-HR1TOCA1 interaction.	DISCUSS
+0	5	Cdc42	protein	Cdc42-HR1TOCA1 binding would then be favorable, as long as coincident activation of Cdc42 had occurred, leading to stabilization of TOCA1 at the membrane and downstream activation of N-WASP.	DISCUSS
+6	9	HR1	structure_element	Cdc42-HR1TOCA1 binding would then be favorable, as long as coincident activation of Cdc42 had occurred, leading to stabilization of TOCA1 at the membrane and downstream activation of N-WASP.	DISCUSS
+9	14	TOCA1	protein	Cdc42-HR1TOCA1 binding would then be favorable, as long as coincident activation of Cdc42 had occurred, leading to stabilization of TOCA1 at the membrane and downstream activation of N-WASP.	DISCUSS
+84	89	Cdc42	protein	Cdc42-HR1TOCA1 binding would then be favorable, as long as coincident activation of Cdc42 had occurred, leading to stabilization of TOCA1 at the membrane and downstream activation of N-WASP.	DISCUSS
+132	137	TOCA1	protein	Cdc42-HR1TOCA1 binding would then be favorable, as long as coincident activation of Cdc42 had occurred, leading to stabilization of TOCA1 at the membrane and downstream activation of N-WASP.	DISCUSS
+183	189	N-WASP	protein	Cdc42-HR1TOCA1 binding would then be favorable, as long as coincident activation of Cdc42 had occurred, leading to stabilization of TOCA1 at the membrane and downstream activation of N-WASP.	DISCUSS
+28	32	WASP	protein_type	It has been postulated that WASP and N-WASP exist in equilibrium between folded (inactive) and unfolded (active) forms, and the affinity of Cdc42 for the unfolded WASP proteins is significantly enhanced.	DISCUSS
+37	43	N-WASP	protein	It has been postulated that WASP and N-WASP exist in equilibrium between folded (inactive) and unfolded (active) forms, and the affinity of Cdc42 for the unfolded WASP proteins is significantly enhanced.	DISCUSS
+73	79	folded	protein_state	It has been postulated that WASP and N-WASP exist in equilibrium between folded (inactive) and unfolded (active) forms, and the affinity of Cdc42 for the unfolded WASP proteins is significantly enhanced.	DISCUSS
+81	89	inactive	protein_state	It has been postulated that WASP and N-WASP exist in equilibrium between folded (inactive) and unfolded (active) forms, and the affinity of Cdc42 for the unfolded WASP proteins is significantly enhanced.	DISCUSS
+95	103	unfolded	protein_state	It has been postulated that WASP and N-WASP exist in equilibrium between folded (inactive) and unfolded (active) forms, and the affinity of Cdc42 for the unfolded WASP proteins is significantly enhanced.	DISCUSS
+105	111	active	protein_state	It has been postulated that WASP and N-WASP exist in equilibrium between folded (inactive) and unfolded (active) forms, and the affinity of Cdc42 for the unfolded WASP proteins is significantly enhanced.	DISCUSS
+128	136	affinity	evidence	It has been postulated that WASP and N-WASP exist in equilibrium between folded (inactive) and unfolded (active) forms, and the affinity of Cdc42 for the unfolded WASP proteins is significantly enhanced.	DISCUSS
+140	145	Cdc42	protein	It has been postulated that WASP and N-WASP exist in equilibrium between folded (inactive) and unfolded (active) forms, and the affinity of Cdc42 for the unfolded WASP proteins is significantly enhanced.	DISCUSS
+154	162	unfolded	protein_state	It has been postulated that WASP and N-WASP exist in equilibrium between folded (inactive) and unfolded (active) forms, and the affinity of Cdc42 for the unfolded WASP proteins is significantly enhanced.	DISCUSS
+163	167	WASP	protein_type	It has been postulated that WASP and N-WASP exist in equilibrium between folded (inactive) and unfolded (active) forms, and the affinity of Cdc42 for the unfolded WASP proteins is significantly enhanced.	DISCUSS
+4	12	unfolded	protein_state	The unfolded, high affinity state of WASP is represented by a short peptide, the GBD, which binds with a low nanomolar affinity to Cdc42.	DISCUSS
+37	41	WASP	protein_type	The unfolded, high affinity state of WASP is represented by a short peptide, the GBD, which binds with a low nanomolar affinity to Cdc42.	DISCUSS
+68	75	peptide	chemical	The unfolded, high affinity state of WASP is represented by a short peptide, the GBD, which binds with a low nanomolar affinity to Cdc42.	DISCUSS
+81	84	GBD	structure_element	The unfolded, high affinity state of WASP is represented by a short peptide, the GBD, which binds with a low nanomolar affinity to Cdc42.	DISCUSS
+131	136	Cdc42	protein	The unfolded, high affinity state of WASP is represented by a short peptide, the GBD, which binds with a low nanomolar affinity to Cdc42.	DISCUSS
+38	46	affinity	evidence	In contrast, the best estimate of the affinity of full-length WASP for Cdc42 is low micromolar.	DISCUSS
+50	61	full-length	protein_state	In contrast, the best estimate of the affinity of full-length WASP for Cdc42 is low micromolar.	DISCUSS
+62	66	WASP	protein_type	In contrast, the best estimate of the affinity of full-length WASP for Cdc42 is low micromolar.	DISCUSS
+71	76	Cdc42	protein	In contrast, the best estimate of the affinity of full-length WASP for Cdc42 is low micromolar.	DISCUSS
+7	15	inactive	protein_state	In the inactive state of WASP, the actin- and Arp2/3-binding VCA domain contacts the GBD, competing for Cdc42 binding.	DISCUSS
+25	29	WASP	protein_type	In the inactive state of WASP, the actin- and Arp2/3-binding VCA domain contacts the GBD, competing for Cdc42 binding.	DISCUSS
+46	52	Arp2/3	complex_assembly	In the inactive state of WASP, the actin- and Arp2/3-binding VCA domain contacts the GBD, competing for Cdc42 binding.	DISCUSS
+61	64	VCA	structure_element	In the inactive state of WASP, the actin- and Arp2/3-binding VCA domain contacts the GBD, competing for Cdc42 binding.	DISCUSS
+85	88	GBD	structure_element	In the inactive state of WASP, the actin- and Arp2/3-binding VCA domain contacts the GBD, competing for Cdc42 binding.	DISCUSS
+104	109	Cdc42	protein	In the inactive state of WASP, the actin- and Arp2/3-binding VCA domain contacts the GBD, competing for Cdc42 binding.	DISCUSS
+21	26	Cdc42	protein	The high affinity of Cdc42 for the unfolded, active form pushes the equilibrium in favor of (N-)WASP activation.	DISCUSS
+35	43	unfolded	protein_state	The high affinity of Cdc42 for the unfolded, active form pushes the equilibrium in favor of (N-)WASP activation.	DISCUSS
+45	51	active	protein_state	The high affinity of Cdc42 for the unfolded, active form pushes the equilibrium in favor of (N-)WASP activation.	DISCUSS
+92	100	(N-)WASP	protein	The high affinity of Cdc42 for the unfolded, active form pushes the equilibrium in favor of (N-)WASP activation.	DISCUSS
+11	20	PI(4,5)P2	chemical	Binding of PI(4,5)P2 to the basic region just N-terminal to the GBD further favors the active conformation.	DISCUSS
+64	67	GBD	structure_element	Binding of PI(4,5)P2 to the basic region just N-terminal to the GBD further favors the active conformation.	DISCUSS
+87	93	active	protein_state	Binding of PI(4,5)P2 to the basic region just N-terminal to the GBD further favors the active conformation.	DISCUSS
+69	89	WASP/N-WASP proteins	protein_type	A substantial body of data has illuminated the complex regulation of WASP/N-WASP proteins, and current evidence suggests that these allosteric activation mechanisms and oligomerization combine to regulate WASP activity, allowing the synchronization and integration of multiple potential activation signals (reviewed in Ref.).	DISCUSS
+205	209	WASP	protein_type	A substantial body of data has illuminated the complex regulation of WASP/N-WASP proteins, and current evidence suggests that these allosteric activation mechanisms and oligomerization combine to regulate WASP activity, allowing the synchronization and integration of multiple potential activation signals (reviewed in Ref.).	DISCUSS
+17	22	TOCA1	protein	We envisage that TOCA1 is first recruited to the appropriate membrane in response to PI(4,5)P2 via its F-BAR domain, where the local increase in concentration favors F-BAR-mediated dimerization of TOCA1.	DISCUSS
+85	94	PI(4,5)P2	chemical	We envisage that TOCA1 is first recruited to the appropriate membrane in response to PI(4,5)P2 via its F-BAR domain, where the local increase in concentration favors F-BAR-mediated dimerization of TOCA1.	DISCUSS
+103	108	F-BAR	structure_element	We envisage that TOCA1 is first recruited to the appropriate membrane in response to PI(4,5)P2 via its F-BAR domain, where the local increase in concentration favors F-BAR-mediated dimerization of TOCA1.	DISCUSS
+166	171	F-BAR	structure_element	We envisage that TOCA1 is first recruited to the appropriate membrane in response to PI(4,5)P2 via its F-BAR domain, where the local increase in concentration favors F-BAR-mediated dimerization of TOCA1.	DISCUSS
+181	186	dimer	oligomeric_state	We envisage that TOCA1 is first recruited to the appropriate membrane in response to PI(4,5)P2 via its F-BAR domain, where the local increase in concentration favors F-BAR-mediated dimerization of TOCA1.	DISCUSS
+197	202	TOCA1	protein	We envisage that TOCA1 is first recruited to the appropriate membrane in response to PI(4,5)P2 via its F-BAR domain, where the local increase in concentration favors F-BAR-mediated dimerization of TOCA1.	DISCUSS
+0	5	Cdc42	protein	Cdc42 is activated in response to co-incident signals and can then bind to TOCA1, further stabilizing TOCA1 at the membrane.	DISCUSS
+75	80	TOCA1	protein	Cdc42 is activated in response to co-incident signals and can then bind to TOCA1, further stabilizing TOCA1 at the membrane.	DISCUSS
+102	107	TOCA1	protein	Cdc42 is activated in response to co-incident signals and can then bind to TOCA1, further stabilizing TOCA1 at the membrane.	DISCUSS
+0	5	TOCA1	protein	TOCA1 can then recruit N-WASP via an interaction between its SH3 domain and the N-WASP proline-rich region.	DISCUSS
+23	29	N-WASP	protein	TOCA1 can then recruit N-WASP via an interaction between its SH3 domain and the N-WASP proline-rich region.	DISCUSS
+61	64	SH3	structure_element	TOCA1 can then recruit N-WASP via an interaction between its SH3 domain and the N-WASP proline-rich region.	DISCUSS
+80	86	N-WASP	protein	TOCA1 can then recruit N-WASP via an interaction between its SH3 domain and the N-WASP proline-rich region.	DISCUSS
+87	106	proline-rich region	structure_element	TOCA1 can then recruit N-WASP via an interaction between its SH3 domain and the N-WASP proline-rich region.	DISCUSS
+19	25	N-WASP	protein	The recruitment of N-WASP alone and of the N-WASP·WIP complex by TOCA1 and FBP17 has been demonstrated.	DISCUSS
+26	31	alone	protein_state	The recruitment of N-WASP alone and of the N-WASP·WIP complex by TOCA1 and FBP17 has been demonstrated.	DISCUSS
+43	53	N-WASP·WIP	complex_assembly	The recruitment of N-WASP alone and of the N-WASP·WIP complex by TOCA1 and FBP17 has been demonstrated.	DISCUSS
+65	70	TOCA1	protein	The recruitment of N-WASP alone and of the N-WASP·WIP complex by TOCA1 and FBP17 has been demonstrated.	DISCUSS
+75	80	FBP17	protein	The recruitment of N-WASP alone and of the N-WASP·WIP complex by TOCA1 and FBP17 has been demonstrated.	DISCUSS
+0	3	WIP	protein	WIP inhibits the activation of N-WASP by Cdc42, an effect that is reversed by TOCA1.	DISCUSS
+31	37	N-WASP	protein	WIP inhibits the activation of N-WASP by Cdc42, an effect that is reversed by TOCA1.	DISCUSS
+41	46	Cdc42	protein	WIP inhibits the activation of N-WASP by Cdc42, an effect that is reversed by TOCA1.	DISCUSS
+78	83	TOCA1	protein	WIP inhibits the activation of N-WASP by Cdc42, an effect that is reversed by TOCA1.	DISCUSS
+35	38	WIP	protein	It may therefore be envisaged that WIP and TOCA1 exert opposing allosteric effects on N-WASP, with TOCA1 favoring the unfolded, active conformation of N-WASP and increasing its affinity for Cdc42.	DISCUSS
+43	48	TOCA1	protein	It may therefore be envisaged that WIP and TOCA1 exert opposing allosteric effects on N-WASP, with TOCA1 favoring the unfolded, active conformation of N-WASP and increasing its affinity for Cdc42.	DISCUSS
+86	92	N-WASP	protein	It may therefore be envisaged that WIP and TOCA1 exert opposing allosteric effects on N-WASP, with TOCA1 favoring the unfolded, active conformation of N-WASP and increasing its affinity for Cdc42.	DISCUSS
+99	104	TOCA1	protein	It may therefore be envisaged that WIP and TOCA1 exert opposing allosteric effects on N-WASP, with TOCA1 favoring the unfolded, active conformation of N-WASP and increasing its affinity for Cdc42.	DISCUSS
+118	126	unfolded	protein_state	It may therefore be envisaged that WIP and TOCA1 exert opposing allosteric effects on N-WASP, with TOCA1 favoring the unfolded, active conformation of N-WASP and increasing its affinity for Cdc42.	DISCUSS
+128	134	active	protein_state	It may therefore be envisaged that WIP and TOCA1 exert opposing allosteric effects on N-WASP, with TOCA1 favoring the unfolded, active conformation of N-WASP and increasing its affinity for Cdc42.	DISCUSS
+151	157	N-WASP	protein	It may therefore be envisaged that WIP and TOCA1 exert opposing allosteric effects on N-WASP, with TOCA1 favoring the unfolded, active conformation of N-WASP and increasing its affinity for Cdc42.	DISCUSS
+190	195	Cdc42	protein	It may therefore be envisaged that WIP and TOCA1 exert opposing allosteric effects on N-WASP, with TOCA1 favoring the unfolded, active conformation of N-WASP and increasing its affinity for Cdc42.	DISCUSS
+0	5	TOCA1	protein	TOCA1 may also activate N-WASP by effective oligomerization because clustering of TOCA1 at the membrane following coincident interactions with PI(4,5)P2 and Cdc42 would in turn lead to clustering of N-WASP, in addition to pushing the equilibrium toward the unfolded, active state.	DISCUSS
+24	30	N-WASP	protein	TOCA1 may also activate N-WASP by effective oligomerization because clustering of TOCA1 at the membrane following coincident interactions with PI(4,5)P2 and Cdc42 would in turn lead to clustering of N-WASP, in addition to pushing the equilibrium toward the unfolded, active state.	DISCUSS
+82	87	TOCA1	protein	TOCA1 may also activate N-WASP by effective oligomerization because clustering of TOCA1 at the membrane following coincident interactions with PI(4,5)P2 and Cdc42 would in turn lead to clustering of N-WASP, in addition to pushing the equilibrium toward the unfolded, active state.	DISCUSS
+143	152	PI(4,5)P2	chemical	TOCA1 may also activate N-WASP by effective oligomerization because clustering of TOCA1 at the membrane following coincident interactions with PI(4,5)P2 and Cdc42 would in turn lead to clustering of N-WASP, in addition to pushing the equilibrium toward the unfolded, active state.	DISCUSS
+157	162	Cdc42	protein	TOCA1 may also activate N-WASP by effective oligomerization because clustering of TOCA1 at the membrane following coincident interactions with PI(4,5)P2 and Cdc42 would in turn lead to clustering of N-WASP, in addition to pushing the equilibrium toward the unfolded, active state.	DISCUSS
+199	205	N-WASP	protein	TOCA1 may also activate N-WASP by effective oligomerization because clustering of TOCA1 at the membrane following coincident interactions with PI(4,5)P2 and Cdc42 would in turn lead to clustering of N-WASP, in addition to pushing the equilibrium toward the unfolded, active state.	DISCUSS
+257	265	unfolded	protein_state	TOCA1 may also activate N-WASP by effective oligomerization because clustering of TOCA1 at the membrane following coincident interactions with PI(4,5)P2 and Cdc42 would in turn lead to clustering of N-WASP, in addition to pushing the equilibrium toward the unfolded, active state.	DISCUSS
+267	273	active	protein_state	TOCA1 may also activate N-WASP by effective oligomerization because clustering of TOCA1 at the membrane following coincident interactions with PI(4,5)P2 and Cdc42 would in turn lead to clustering of N-WASP, in addition to pushing the equilibrium toward the unfolded, active state.	DISCUSS
+23	34	full-length	protein_state	In a cellular context, full-length TOCA1 and N-WASP are likely to have similar affinities for active Cdc42, but in the unfolded, active conformation, the affinity of N-WASP for Cdc42 dramatically increases.	DISCUSS
+35	40	TOCA1	protein	In a cellular context, full-length TOCA1 and N-WASP are likely to have similar affinities for active Cdc42, but in the unfolded, active conformation, the affinity of N-WASP for Cdc42 dramatically increases.	DISCUSS
+45	51	N-WASP	protein	In a cellular context, full-length TOCA1 and N-WASP are likely to have similar affinities for active Cdc42, but in the unfolded, active conformation, the affinity of N-WASP for Cdc42 dramatically increases.	DISCUSS
+79	89	affinities	evidence	In a cellular context, full-length TOCA1 and N-WASP are likely to have similar affinities for active Cdc42, but in the unfolded, active conformation, the affinity of N-WASP for Cdc42 dramatically increases.	DISCUSS
+94	100	active	protein_state	In a cellular context, full-length TOCA1 and N-WASP are likely to have similar affinities for active Cdc42, but in the unfolded, active conformation, the affinity of N-WASP for Cdc42 dramatically increases.	DISCUSS
+101	106	Cdc42	protein	In a cellular context, full-length TOCA1 and N-WASP are likely to have similar affinities for active Cdc42, but in the unfolded, active conformation, the affinity of N-WASP for Cdc42 dramatically increases.	DISCUSS
+119	127	unfolded	protein_state	In a cellular context, full-length TOCA1 and N-WASP are likely to have similar affinities for active Cdc42, but in the unfolded, active conformation, the affinity of N-WASP for Cdc42 dramatically increases.	DISCUSS
+129	135	active	protein_state	In a cellular context, full-length TOCA1 and N-WASP are likely to have similar affinities for active Cdc42, but in the unfolded, active conformation, the affinity of N-WASP for Cdc42 dramatically increases.	DISCUSS
+154	162	affinity	evidence	In a cellular context, full-length TOCA1 and N-WASP are likely to have similar affinities for active Cdc42, but in the unfolded, active conformation, the affinity of N-WASP for Cdc42 dramatically increases.	DISCUSS
+166	172	N-WASP	protein	In a cellular context, full-length TOCA1 and N-WASP are likely to have similar affinities for active Cdc42, but in the unfolded, active conformation, the affinity of N-WASP for Cdc42 dramatically increases.	DISCUSS
+177	182	Cdc42	protein	In a cellular context, full-length TOCA1 and N-WASP are likely to have similar affinities for active Cdc42, but in the unfolded, active conformation, the affinity of N-WASP for Cdc42 dramatically increases.	DISCUSS
+4	16	binding data	evidence	Our binding data suggest that TOCA1 HR1 binding is not allosterically regulated, and our NMR data, along with the high stability of TOCA1 HR1, suggest that there is no widespread conformational change in the presence of Cdc42.	DISCUSS
+30	35	TOCA1	protein	Our binding data suggest that TOCA1 HR1 binding is not allosterically regulated, and our NMR data, along with the high stability of TOCA1 HR1, suggest that there is no widespread conformational change in the presence of Cdc42.	DISCUSS
+36	39	HR1	structure_element	Our binding data suggest that TOCA1 HR1 binding is not allosterically regulated, and our NMR data, along with the high stability of TOCA1 HR1, suggest that there is no widespread conformational change in the presence of Cdc42.	DISCUSS
+89	92	NMR	experimental_method	Our binding data suggest that TOCA1 HR1 binding is not allosterically regulated, and our NMR data, along with the high stability of TOCA1 HR1, suggest that there is no widespread conformational change in the presence of Cdc42.	DISCUSS
+119	128	stability	protein_state	Our binding data suggest that TOCA1 HR1 binding is not allosterically regulated, and our NMR data, along with the high stability of TOCA1 HR1, suggest that there is no widespread conformational change in the presence of Cdc42.	DISCUSS
+132	137	TOCA1	protein	Our binding data suggest that TOCA1 HR1 binding is not allosterically regulated, and our NMR data, along with the high stability of TOCA1 HR1, suggest that there is no widespread conformational change in the presence of Cdc42.	DISCUSS
+138	141	HR1	structure_element	Our binding data suggest that TOCA1 HR1 binding is not allosterically regulated, and our NMR data, along with the high stability of TOCA1 HR1, suggest that there is no widespread conformational change in the presence of Cdc42.	DISCUSS
+208	219	presence of	protein_state	Our binding data suggest that TOCA1 HR1 binding is not allosterically regulated, and our NMR data, along with the high stability of TOCA1 HR1, suggest that there is no widespread conformational change in the presence of Cdc42.	DISCUSS
+220	225	Cdc42	protein	Our binding data suggest that TOCA1 HR1 binding is not allosterically regulated, and our NMR data, along with the high stability of TOCA1 HR1, suggest that there is no widespread conformational change in the presence of Cdc42.	DISCUSS
+3	14	full-length	protein_state	As full-length TOCA1 and the isolated HR1 domain bind Cdc42 with similar affinities, the N-WASP-Cdc42 interaction will be favored because the N-WASP GBD can easily outcompete the TOCA1 HR1 for Cdc42.	DISCUSS
+15	20	TOCA1	protein	As full-length TOCA1 and the isolated HR1 domain bind Cdc42 with similar affinities, the N-WASP-Cdc42 interaction will be favored because the N-WASP GBD can easily outcompete the TOCA1 HR1 for Cdc42.	DISCUSS
+29	37	isolated	protein_state	As full-length TOCA1 and the isolated HR1 domain bind Cdc42 with similar affinities, the N-WASP-Cdc42 interaction will be favored because the N-WASP GBD can easily outcompete the TOCA1 HR1 for Cdc42.	DISCUSS
+38	41	HR1	structure_element	As full-length TOCA1 and the isolated HR1 domain bind Cdc42 with similar affinities, the N-WASP-Cdc42 interaction will be favored because the N-WASP GBD can easily outcompete the TOCA1 HR1 for Cdc42.	DISCUSS
+54	59	Cdc42	protein	As full-length TOCA1 and the isolated HR1 domain bind Cdc42 with similar affinities, the N-WASP-Cdc42 interaction will be favored because the N-WASP GBD can easily outcompete the TOCA1 HR1 for Cdc42.	DISCUSS
+89	95	N-WASP	protein	As full-length TOCA1 and the isolated HR1 domain bind Cdc42 with similar affinities, the N-WASP-Cdc42 interaction will be favored because the N-WASP GBD can easily outcompete the TOCA1 HR1 for Cdc42.	DISCUSS
+96	101	Cdc42	protein	As full-length TOCA1 and the isolated HR1 domain bind Cdc42 with similar affinities, the N-WASP-Cdc42 interaction will be favored because the N-WASP GBD can easily outcompete the TOCA1 HR1 for Cdc42.	DISCUSS
+142	148	N-WASP	protein	As full-length TOCA1 and the isolated HR1 domain bind Cdc42 with similar affinities, the N-WASP-Cdc42 interaction will be favored because the N-WASP GBD can easily outcompete the TOCA1 HR1 for Cdc42.	DISCUSS
+149	152	GBD	structure_element	As full-length TOCA1 and the isolated HR1 domain bind Cdc42 with similar affinities, the N-WASP-Cdc42 interaction will be favored because the N-WASP GBD can easily outcompete the TOCA1 HR1 for Cdc42.	DISCUSS
+179	184	TOCA1	protein	As full-length TOCA1 and the isolated HR1 domain bind Cdc42 with similar affinities, the N-WASP-Cdc42 interaction will be favored because the N-WASP GBD can easily outcompete the TOCA1 HR1 for Cdc42.	DISCUSS
+185	188	HR1	structure_element	As full-length TOCA1 and the isolated HR1 domain bind Cdc42 with similar affinities, the N-WASP-Cdc42 interaction will be favored because the N-WASP GBD can easily outcompete the TOCA1 HR1 for Cdc42.	DISCUSS
+193	198	Cdc42	protein	As full-length TOCA1 and the isolated HR1 domain bind Cdc42 with similar affinities, the N-WASP-Cdc42 interaction will be favored because the N-WASP GBD can easily outcompete the TOCA1 HR1 for Cdc42.	DISCUSS
+42	51	PI(4,5)P2	chemical	A combination of allosteric activation by PI(4,5)P2, activated Cdc42 and TOCA1, and oligomeric activation implemented by TOCA1 would lead to full activation of N-WASP and downstream actin polymerization.	DISCUSS
+53	62	activated	protein_state	A combination of allosteric activation by PI(4,5)P2, activated Cdc42 and TOCA1, and oligomeric activation implemented by TOCA1 would lead to full activation of N-WASP and downstream actin polymerization.	DISCUSS
+63	68	Cdc42	protein	A combination of allosteric activation by PI(4,5)P2, activated Cdc42 and TOCA1, and oligomeric activation implemented by TOCA1 would lead to full activation of N-WASP and downstream actin polymerization.	DISCUSS
+73	78	TOCA1	protein	A combination of allosteric activation by PI(4,5)P2, activated Cdc42 and TOCA1, and oligomeric activation implemented by TOCA1 would lead to full activation of N-WASP and downstream actin polymerization.	DISCUSS
+121	126	TOCA1	protein	A combination of allosteric activation by PI(4,5)P2, activated Cdc42 and TOCA1, and oligomeric activation implemented by TOCA1 would lead to full activation of N-WASP and downstream actin polymerization.	DISCUSS
+141	156	full activation	protein_state	A combination of allosteric activation by PI(4,5)P2, activated Cdc42 and TOCA1, and oligomeric activation implemented by TOCA1 would lead to full activation of N-WASP and downstream actin polymerization.	DISCUSS
+160	166	N-WASP	protein	A combination of allosteric activation by PI(4,5)P2, activated Cdc42 and TOCA1, and oligomeric activation implemented by TOCA1 would lead to full activation of N-WASP and downstream actin polymerization.	DISCUSS
+99	103	WASP	protein	In such an array of molecules localized to a discrete region of the membrane, it is plausible that WASP could bind to a second Cdc42 molecule rather than displacing TOCA1 from its cognate Cdc42.	DISCUSS
+127	132	Cdc42	protein	In such an array of molecules localized to a discrete region of the membrane, it is plausible that WASP could bind to a second Cdc42 molecule rather than displacing TOCA1 from its cognate Cdc42.	DISCUSS
+165	170	TOCA1	protein	In such an array of molecules localized to a discrete region of the membrane, it is plausible that WASP could bind to a second Cdc42 molecule rather than displacing TOCA1 from its cognate Cdc42.	DISCUSS
+188	193	Cdc42	protein	In such an array of molecules localized to a discrete region of the membrane, it is plausible that WASP could bind to a second Cdc42 molecule rather than displacing TOCA1 from its cognate Cdc42.	DISCUSS
+4	7	NMR	experimental_method	Our NMR and affinity data, however, are consistent with displacement of the TOCA1 HR1 by the N-WASP GBD.	DISCUSS
+12	25	affinity data	evidence	Our NMR and affinity data, however, are consistent with displacement of the TOCA1 HR1 by the N-WASP GBD.	DISCUSS
+76	81	TOCA1	protein	Our NMR and affinity data, however, are consistent with displacement of the TOCA1 HR1 by the N-WASP GBD.	DISCUSS
+82	85	HR1	structure_element	Our NMR and affinity data, however, are consistent with displacement of the TOCA1 HR1 by the N-WASP GBD.	DISCUSS
+93	99	N-WASP	protein	Our NMR and affinity data, however, are consistent with displacement of the TOCA1 HR1 by the N-WASP GBD.	DISCUSS
+100	103	GBD	structure_element	Our NMR and affinity data, however, are consistent with displacement of the TOCA1 HR1 by the N-WASP GBD.	DISCUSS
+13	18	TOCA1	protein	Furthermore, TOCA1 is required for Cdc42-mediated activation of N-WASP·WIP, implying that it may not be possible for Cdc42 to bind and activate N-WASP prior to TOCA1-Cdc42 binding.	DISCUSS
+35	40	Cdc42	protein	Furthermore, TOCA1 is required for Cdc42-mediated activation of N-WASP·WIP, implying that it may not be possible for Cdc42 to bind and activate N-WASP prior to TOCA1-Cdc42 binding.	DISCUSS
+64	74	N-WASP·WIP	complex_assembly	Furthermore, TOCA1 is required for Cdc42-mediated activation of N-WASP·WIP, implying that it may not be possible for Cdc42 to bind and activate N-WASP prior to TOCA1-Cdc42 binding.	DISCUSS
+117	122	Cdc42	protein	Furthermore, TOCA1 is required for Cdc42-mediated activation of N-WASP·WIP, implying that it may not be possible for Cdc42 to bind and activate N-WASP prior to TOCA1-Cdc42 binding.	DISCUSS
+144	150	N-WASP	protein	Furthermore, TOCA1 is required for Cdc42-mediated activation of N-WASP·WIP, implying that it may not be possible for Cdc42 to bind and activate N-WASP prior to TOCA1-Cdc42 binding.	DISCUSS
+160	165	TOCA1	protein	Furthermore, TOCA1 is required for Cdc42-mediated activation of N-WASP·WIP, implying that it may not be possible for Cdc42 to bind and activate N-WASP prior to TOCA1-Cdc42 binding.	DISCUSS
+166	171	Cdc42	protein	Furthermore, TOCA1 is required for Cdc42-mediated activation of N-WASP·WIP, implying that it may not be possible for Cdc42 to bind and activate N-WASP prior to TOCA1-Cdc42 binding.	DISCUSS
+18	27	MGD → IST	mutant	The commonly used MGD → IST (Cdc42-binding deficient) mutant of TOCA1 has a reduced ability to activate the N-WASP·WIP complex, further indicating the importance of the Cdc42-HR1TOCA1 interaction prior to downstream activation of N-WASP.	DISCUSS
+29	52	Cdc42-binding deficient	protein_state	The commonly used MGD → IST (Cdc42-binding deficient) mutant of TOCA1 has a reduced ability to activate the N-WASP·WIP complex, further indicating the importance of the Cdc42-HR1TOCA1 interaction prior to downstream activation of N-WASP.	DISCUSS
+64	69	TOCA1	protein	The commonly used MGD → IST (Cdc42-binding deficient) mutant of TOCA1 has a reduced ability to activate the N-WASP·WIP complex, further indicating the importance of the Cdc42-HR1TOCA1 interaction prior to downstream activation of N-WASP.	DISCUSS
+108	118	N-WASP·WIP	complex_assembly	The commonly used MGD → IST (Cdc42-binding deficient) mutant of TOCA1 has a reduced ability to activate the N-WASP·WIP complex, further indicating the importance of the Cdc42-HR1TOCA1 interaction prior to downstream activation of N-WASP.	DISCUSS
+169	174	Cdc42	protein	The commonly used MGD → IST (Cdc42-binding deficient) mutant of TOCA1 has a reduced ability to activate the N-WASP·WIP complex, further indicating the importance of the Cdc42-HR1TOCA1 interaction prior to downstream activation of N-WASP.	DISCUSS
+175	178	HR1	structure_element	The commonly used MGD → IST (Cdc42-binding deficient) mutant of TOCA1 has a reduced ability to activate the N-WASP·WIP complex, further indicating the importance of the Cdc42-HR1TOCA1 interaction prior to downstream activation of N-WASP.	DISCUSS
+178	183	TOCA1	protein	The commonly used MGD → IST (Cdc42-binding deficient) mutant of TOCA1 has a reduced ability to activate the N-WASP·WIP complex, further indicating the importance of the Cdc42-HR1TOCA1 interaction prior to downstream activation of N-WASP.	DISCUSS
+230	236	N-WASP	protein	The commonly used MGD → IST (Cdc42-binding deficient) mutant of TOCA1 has a reduced ability to activate the N-WASP·WIP complex, further indicating the importance of the Cdc42-HR1TOCA1 interaction prior to downstream activation of N-WASP.	DISCUSS
+65	70	TOCA1	protein	In light of this, we favor an “effector handover” scheme whereby TOCA1 interacts with Cdc42 prior to N-WASP activation, after which N-WASP displaces TOCA1 from its bound Cdc42 in order to be fully activated rather than binding a second Cdc42 molecule.	DISCUSS
+86	91	Cdc42	protein	In light of this, we favor an “effector handover” scheme whereby TOCA1 interacts with Cdc42 prior to N-WASP activation, after which N-WASP displaces TOCA1 from its bound Cdc42 in order to be fully activated rather than binding a second Cdc42 molecule.	DISCUSS
+101	107	N-WASP	protein	In light of this, we favor an “effector handover” scheme whereby TOCA1 interacts with Cdc42 prior to N-WASP activation, after which N-WASP displaces TOCA1 from its bound Cdc42 in order to be fully activated rather than binding a second Cdc42 molecule.	DISCUSS
+132	138	N-WASP	protein	In light of this, we favor an “effector handover” scheme whereby TOCA1 interacts with Cdc42 prior to N-WASP activation, after which N-WASP displaces TOCA1 from its bound Cdc42 in order to be fully activated rather than binding a second Cdc42 molecule.	DISCUSS
+149	154	TOCA1	protein	In light of this, we favor an “effector handover” scheme whereby TOCA1 interacts with Cdc42 prior to N-WASP activation, after which N-WASP displaces TOCA1 from its bound Cdc42 in order to be fully activated rather than binding a second Cdc42 molecule.	DISCUSS
+164	169	bound	protein_state	In light of this, we favor an “effector handover” scheme whereby TOCA1 interacts with Cdc42 prior to N-WASP activation, after which N-WASP displaces TOCA1 from its bound Cdc42 in order to be fully activated rather than binding a second Cdc42 molecule.	DISCUSS
+170	175	Cdc42	protein	In light of this, we favor an “effector handover” scheme whereby TOCA1 interacts with Cdc42 prior to N-WASP activation, after which N-WASP displaces TOCA1 from its bound Cdc42 in order to be fully activated rather than binding a second Cdc42 molecule.	DISCUSS
+191	206	fully activated	protein_state	In light of this, we favor an “effector handover” scheme whereby TOCA1 interacts with Cdc42 prior to N-WASP activation, after which N-WASP displaces TOCA1 from its bound Cdc42 in order to be fully activated rather than binding a second Cdc42 molecule.	DISCUSS
+236	241	Cdc42	protein	In light of this, we favor an “effector handover” scheme whereby TOCA1 interacts with Cdc42 prior to N-WASP activation, after which N-WASP displaces TOCA1 from its bound Cdc42 in order to be fully activated rather than binding a second Cdc42 molecule.	DISCUSS
+17	22	TOCA1	protein	Potentially, the TOCA1-Cdc42 interaction functions to position N-WASP and Cdc42 such that they are poised to interact with high affinity.	DISCUSS
+23	28	Cdc42	protein	Potentially, the TOCA1-Cdc42 interaction functions to position N-WASP and Cdc42 such that they are poised to interact with high affinity.	DISCUSS
+63	69	N-WASP	protein	Potentially, the TOCA1-Cdc42 interaction functions to position N-WASP and Cdc42 such that they are poised to interact with high affinity.	DISCUSS
+74	79	Cdc42	protein	Potentially, the TOCA1-Cdc42 interaction functions to position N-WASP and Cdc42 such that they are poised to interact with high affinity.	DISCUSS
+27	32	TOCA1	protein	The concomitant release of TOCA1 from Cdc42 while still bound to N-WASP presumably enhances the ability of TOCA1 to further activate N-WASP·WIP-induced actin polymerization.	DISCUSS
+38	43	Cdc42	protein	The concomitant release of TOCA1 from Cdc42 while still bound to N-WASP presumably enhances the ability of TOCA1 to further activate N-WASP·WIP-induced actin polymerization.	DISCUSS
+56	64	bound to	protein_state	The concomitant release of TOCA1 from Cdc42 while still bound to N-WASP presumably enhances the ability of TOCA1 to further activate N-WASP·WIP-induced actin polymerization.	DISCUSS
+65	71	N-WASP	protein	The concomitant release of TOCA1 from Cdc42 while still bound to N-WASP presumably enhances the ability of TOCA1 to further activate N-WASP·WIP-induced actin polymerization.	DISCUSS
+107	112	TOCA1	protein	The concomitant release of TOCA1 from Cdc42 while still bound to N-WASP presumably enhances the ability of TOCA1 to further activate N-WASP·WIP-induced actin polymerization.	DISCUSS
+133	143	N-WASP·WIP	complex_assembly	The concomitant release of TOCA1 from Cdc42 while still bound to N-WASP presumably enhances the ability of TOCA1 to further activate N-WASP·WIP-induced actin polymerization.	DISCUSS
+60	66	N-WASP	protein	There is an advantage to such an effector handover, in that N-WASP would only be robustly recruited when F-BAR domains are already present.	DISCUSS
+105	110	F-BAR	structure_element	There is an advantage to such an effector handover, in that N-WASP would only be robustly recruited when F-BAR domains are already present.	DISCUSS
+47	52	F-BAR	structure_element	Hence, actin polymerization cannot occur until F-BAR domains are poised for membrane distortion.	DISCUSS
+17	31	Cdc42·HR1TOCA1	complex_assembly	Our model of the Cdc42·HR1TOCA1 complex indicates a mechanism by which such a handover could take place (Fig. 9) because it shows that the effector binding sites only partially overlap on Cdc42.	DISCUSS
+139	161	effector binding sites	site	Our model of the Cdc42·HR1TOCA1 complex indicates a mechanism by which such a handover could take place (Fig. 9) because it shows that the effector binding sites only partially overlap on Cdc42.	DISCUSS
+188	193	Cdc42	protein	Our model of the Cdc42·HR1TOCA1 complex indicates a mechanism by which such a handover could take place (Fig. 9) because it shows that the effector binding sites only partially overlap on Cdc42.	DISCUSS
+4	10	lysine	residue_name	The lysine residues thought to be involved in an electrostatic steering mechanism in WASP-Cdc42 binding are conserved in N-WASP and would be able to interact with Cdc42 even when the TOCA1 HR1 domain is already bound.	DISCUSS
+85	89	WASP	protein	The lysine residues thought to be involved in an electrostatic steering mechanism in WASP-Cdc42 binding are conserved in N-WASP and would be able to interact with Cdc42 even when the TOCA1 HR1 domain is already bound.	DISCUSS
+90	95	Cdc42	protein	The lysine residues thought to be involved in an electrostatic steering mechanism in WASP-Cdc42 binding are conserved in N-WASP and would be able to interact with Cdc42 even when the TOCA1 HR1 domain is already bound.	DISCUSS
+121	127	N-WASP	protein	The lysine residues thought to be involved in an electrostatic steering mechanism in WASP-Cdc42 binding are conserved in N-WASP and would be able to interact with Cdc42 even when the TOCA1 HR1 domain is already bound.	DISCUSS
+163	168	Cdc42	protein	The lysine residues thought to be involved in an electrostatic steering mechanism in WASP-Cdc42 binding are conserved in N-WASP and would be able to interact with Cdc42 even when the TOCA1 HR1 domain is already bound.	DISCUSS
+183	188	TOCA1	protein	The lysine residues thought to be involved in an electrostatic steering mechanism in WASP-Cdc42 binding are conserved in N-WASP and would be able to interact with Cdc42 even when the TOCA1 HR1 domain is already bound.	DISCUSS
+189	192	HR1	structure_element	The lysine residues thought to be involved in an electrostatic steering mechanism in WASP-Cdc42 binding are conserved in N-WASP and would be able to interact with Cdc42 even when the TOCA1 HR1 domain is already bound.	DISCUSS
+211	216	bound	protein_state	The lysine residues thought to be involved in an electrostatic steering mechanism in WASP-Cdc42 binding are conserved in N-WASP and would be able to interact with Cdc42 even when the TOCA1 HR1 domain is already bound.	DISCUSS
+83	88	Cdc42	protein	It has been postulated that the initial interactions between this basic region and Cdc42 could stabilize the active conformation of WASP, leading to high affinity binding between the core CRIB and Cdc42.	DISCUSS
+109	115	active	protein_state	It has been postulated that the initial interactions between this basic region and Cdc42 could stabilize the active conformation of WASP, leading to high affinity binding between the core CRIB and Cdc42.	DISCUSS
+132	136	WASP	protein	It has been postulated that the initial interactions between this basic region and Cdc42 could stabilize the active conformation of WASP, leading to high affinity binding between the core CRIB and Cdc42.	DISCUSS
+188	192	CRIB	structure_element	It has been postulated that the initial interactions between this basic region and Cdc42 could stabilize the active conformation of WASP, leading to high affinity binding between the core CRIB and Cdc42.	DISCUSS
+197	202	Cdc42	protein	It has been postulated that the initial interactions between this basic region and Cdc42 could stabilize the active conformation of WASP, leading to high affinity binding between the core CRIB and Cdc42.	DISCUSS
+34	38	CRIB	structure_element	The region C-terminal to the core CRIB, required for maximal affinity binding, would then fully displace the TOCA1 HR1.	DISCUSS
+109	114	TOCA1	protein	The region C-terminal to the core CRIB, required for maximal affinity binding, would then fully displace the TOCA1 HR1.	DISCUSS
+115	118	HR1	structure_element	The region C-terminal to the core CRIB, required for maximal affinity binding, would then fully displace the TOCA1 HR1.	DISCUSS
+42	60	Cdc42·N-WASP·TOCA1	complex_assembly	A simplified model of the early stages of Cdc42·N-WASP·TOCA1-dependent actin polymerization.	FIG
+9	14	TOCA1	protein	 Step 1, TOCA1 is recruited to the membrane via its F-BAR domain and/or Cdc42 interactions.	FIG
+52	57	F-BAR	structure_element	 Step 1, TOCA1 is recruited to the membrane via its F-BAR domain and/or Cdc42 interactions.	FIG
+72	77	Cdc42	protein	 Step 1, TOCA1 is recruited to the membrane via its F-BAR domain and/or Cdc42 interactions.	FIG
+84	91	monomer	oligomeric_state	F-BAR oligomerization is expected to occur following membrane binding, but a single monomer is shown for clarity.	FIG
+8	14	N-WASP	protein	Step 2, N-WASP exists in an inactive, folded conformation.	FIG
+28	36	inactive	protein_state	Step 2, N-WASP exists in an inactive, folded conformation.	FIG
+38	44	folded	protein_state	Step 2, N-WASP exists in an inactive, folded conformation.	FIG
+4	9	TOCA1	protein	The TOCA1 SH3 domain interacts with N-WASP, causing an activatory allosteric effect.	FIG
+10	13	SH3	structure_element	The TOCA1 SH3 domain interacts with N-WASP, causing an activatory allosteric effect.	FIG
+36	42	N-WASP	protein	The TOCA1 SH3 domain interacts with N-WASP, causing an activatory allosteric effect.	FIG
+4	7	HR1	structure_element	The HR1TOCA1-Cdc42 and SH3TOCA1-N-WASP interactions position Cdc42 and N-WASP for binding.	FIG
+23	26	SH3	structure_element	The HR1TOCA1-Cdc42 and SH3TOCA1-N-WASP interactions position Cdc42 and N-WASP for binding.	FIG
+61	66	Cdc42	protein	The HR1TOCA1-Cdc42 and SH3TOCA1-N-WASP interactions position Cdc42 and N-WASP for binding.	FIG
+71	77	N-WASP	protein	The HR1TOCA1-Cdc42 and SH3TOCA1-N-WASP interactions position Cdc42 and N-WASP for binding.	FIG
+43	48	Cdc42	protein	Step 3, electrostatic interactions between Cdc42 and the basic region upstream of the CRIB initiate Cdc42·N-WASP binding.	FIG
+86	90	CRIB	structure_element	Step 3, electrostatic interactions between Cdc42 and the basic region upstream of the CRIB initiate Cdc42·N-WASP binding.	FIG
+100	112	Cdc42·N-WASP	complex_assembly	Step 3, electrostatic interactions between Cdc42 and the basic region upstream of the CRIB initiate Cdc42·N-WASP binding.	FIG
+17	21	CRIB	structure_element	Step 4, the core CRIB binds with high affinity while the region C-terminal to the CRIB displaces the TOCA1 HR1 domain and increases the affinity of the N-WASP-Cdc42 interaction further.	FIG
+82	86	CRIB	structure_element	Step 4, the core CRIB binds with high affinity while the region C-terminal to the CRIB displaces the TOCA1 HR1 domain and increases the affinity of the N-WASP-Cdc42 interaction further.	FIG
+101	106	TOCA1	protein	Step 4, the core CRIB binds with high affinity while the region C-terminal to the CRIB displaces the TOCA1 HR1 domain and increases the affinity of the N-WASP-Cdc42 interaction further.	FIG
+107	110	HR1	structure_element	Step 4, the core CRIB binds with high affinity while the region C-terminal to the CRIB displaces the TOCA1 HR1 domain and increases the affinity of the N-WASP-Cdc42 interaction further.	FIG
+152	158	N-WASP	protein	Step 4, the core CRIB binds with high affinity while the region C-terminal to the CRIB displaces the TOCA1 HR1 domain and increases the affinity of the N-WASP-Cdc42 interaction further.	FIG
+4	7	VCA	structure_element	The VCA domain is released for downstream interactions, and actin polymerization proceeds.	FIG
+5	27	WASP homology 1 domain	structure_element	WH1, WASP homology 1 domain; PP, proline-rich region; VCA, verprolin homology, cofilin homology, acidic region.	FIG
+29	31	PP	structure_element	WH1, WASP homology 1 domain; PP, proline-rich region; VCA, verprolin homology, cofilin homology, acidic region.	FIG
+33	52	proline-rich region	structure_element	WH1, WASP homology 1 domain; PP, proline-rich region; VCA, verprolin homology, cofilin homology, acidic region.	FIG
+54	57	VCA	structure_element	WH1, WASP homology 1 domain; PP, proline-rich region; VCA, verprolin homology, cofilin homology, acidic region.	FIG
+59	110	verprolin homology, cofilin homology, acidic region	structure_element	WH1, WASP homology 1 domain; PP, proline-rich region; VCA, verprolin homology, cofilin homology, acidic region.	FIG
+53	58	TOCA1	protein	In conclusion, the data presented here show that the TOCA1 HR1 domain is sufficient for Cdc42 binding in vitro and that the interaction is of micromolar affinity, lower than that of other G protein-HR1 domain interactions.	DISCUSS
+59	62	HR1	structure_element	In conclusion, the data presented here show that the TOCA1 HR1 domain is sufficient for Cdc42 binding in vitro and that the interaction is of micromolar affinity, lower than that of other G protein-HR1 domain interactions.	DISCUSS
+88	93	Cdc42	protein	In conclusion, the data presented here show that the TOCA1 HR1 domain is sufficient for Cdc42 binding in vitro and that the interaction is of micromolar affinity, lower than that of other G protein-HR1 domain interactions.	DISCUSS
+188	197	G protein	protein_type	In conclusion, the data presented here show that the TOCA1 HR1 domain is sufficient for Cdc42 binding in vitro and that the interaction is of micromolar affinity, lower than that of other G protein-HR1 domain interactions.	DISCUSS
+198	201	HR1	structure_element	In conclusion, the data presented here show that the TOCA1 HR1 domain is sufficient for Cdc42 binding in vitro and that the interaction is of micromolar affinity, lower than that of other G protein-HR1 domain interactions.	DISCUSS
+14	17	HR1	structure_element	The analogous HR1 domains from other TOCA1 family members, FBP17 and CIP4, also exhibit micromolar affinity for Cdc42.	DISCUSS
+37	49	TOCA1 family	protein_type	The analogous HR1 domains from other TOCA1 family members, FBP17 and CIP4, also exhibit micromolar affinity for Cdc42.	DISCUSS
+59	64	FBP17	protein	The analogous HR1 domains from other TOCA1 family members, FBP17 and CIP4, also exhibit micromolar affinity for Cdc42.	DISCUSS
+69	73	CIP4	protein	The analogous HR1 domains from other TOCA1 family members, FBP17 and CIP4, also exhibit micromolar affinity for Cdc42.	DISCUSS
+112	117	Cdc42	protein	The analogous HR1 domains from other TOCA1 family members, FBP17 and CIP4, also exhibit micromolar affinity for Cdc42.	DISCUSS
+15	20	TOCA1	protein	A role for the TOCA1-, FBP17-, and CIP4-Cdc42 interactions in the recruitment of these proteins to the membrane therefore appears unlikely.	DISCUSS
+23	28	FBP17	protein	A role for the TOCA1-, FBP17-, and CIP4-Cdc42 interactions in the recruitment of these proteins to the membrane therefore appears unlikely.	DISCUSS
+35	39	CIP4	protein	A role for the TOCA1-, FBP17-, and CIP4-Cdc42 interactions in the recruitment of these proteins to the membrane therefore appears unlikely.	DISCUSS
+40	45	Cdc42	protein	A role for the TOCA1-, FBP17-, and CIP4-Cdc42 interactions in the recruitment of these proteins to the membrane therefore appears unlikely.	DISCUSS
+62	67	F-BAR	structure_element	Instead, our findings agree with earlier suggestions that the F-BAR domain is responsible for membrane recruitment.	DISCUSS
+16	21	Cdc42	protein	The role of the Cdc42-TOCA1 interaction remains somewhat elusive, but it may serve to position activated Cdc42 and N-WASP to allow full activation of N-WASP and as such serve to couple F-BAR-mediated membrane deformation with N-WASP activation.	DISCUSS
+22	27	TOCA1	protein	The role of the Cdc42-TOCA1 interaction remains somewhat elusive, but it may serve to position activated Cdc42 and N-WASP to allow full activation of N-WASP and as such serve to couple F-BAR-mediated membrane deformation with N-WASP activation.	DISCUSS
+95	104	activated	protein_state	The role of the Cdc42-TOCA1 interaction remains somewhat elusive, but it may serve to position activated Cdc42 and N-WASP to allow full activation of N-WASP and as such serve to couple F-BAR-mediated membrane deformation with N-WASP activation.	DISCUSS
+105	110	Cdc42	protein	The role of the Cdc42-TOCA1 interaction remains somewhat elusive, but it may serve to position activated Cdc42 and N-WASP to allow full activation of N-WASP and as such serve to couple F-BAR-mediated membrane deformation with N-WASP activation.	DISCUSS
+115	121	N-WASP	protein	The role of the Cdc42-TOCA1 interaction remains somewhat elusive, but it may serve to position activated Cdc42 and N-WASP to allow full activation of N-WASP and as such serve to couple F-BAR-mediated membrane deformation with N-WASP activation.	DISCUSS
+131	146	full activation	protein_state	The role of the Cdc42-TOCA1 interaction remains somewhat elusive, but it may serve to position activated Cdc42 and N-WASP to allow full activation of N-WASP and as such serve to couple F-BAR-mediated membrane deformation with N-WASP activation.	DISCUSS
+150	156	N-WASP	protein	The role of the Cdc42-TOCA1 interaction remains somewhat elusive, but it may serve to position activated Cdc42 and N-WASP to allow full activation of N-WASP and as such serve to couple F-BAR-mediated membrane deformation with N-WASP activation.	DISCUSS
+185	190	F-BAR	structure_element	The role of the Cdc42-TOCA1 interaction remains somewhat elusive, but it may serve to position activated Cdc42 and N-WASP to allow full activation of N-WASP and as such serve to couple F-BAR-mediated membrane deformation with N-WASP activation.	DISCUSS
+226	232	N-WASP	protein	The role of the Cdc42-TOCA1 interaction remains somewhat elusive, but it may serve to position activated Cdc42 and N-WASP to allow full activation of N-WASP and as such serve to couple F-BAR-mediated membrane deformation with N-WASP activation.	DISCUSS
+54	58	free	protein_state	We envisage a complex interplay of equilibria between free and bound, active and inactive Cdc42, TOCA family, and WASP family proteins, facilitating a tightly spatially and temporally regulated pathway requiring numerous simultaneous events in order to achieve appropriate and robust activation of the downstream pathway.	DISCUSS
+63	68	bound	protein_state	We envisage a complex interplay of equilibria between free and bound, active and inactive Cdc42, TOCA family, and WASP family proteins, facilitating a tightly spatially and temporally regulated pathway requiring numerous simultaneous events in order to achieve appropriate and robust activation of the downstream pathway.	DISCUSS
+70	76	active	protein_state	We envisage a complex interplay of equilibria between free and bound, active and inactive Cdc42, TOCA family, and WASP family proteins, facilitating a tightly spatially and temporally regulated pathway requiring numerous simultaneous events in order to achieve appropriate and robust activation of the downstream pathway.	DISCUSS
+81	89	inactive	protein_state	We envisage a complex interplay of equilibria between free and bound, active and inactive Cdc42, TOCA family, and WASP family proteins, facilitating a tightly spatially and temporally regulated pathway requiring numerous simultaneous events in order to achieve appropriate and robust activation of the downstream pathway.	DISCUSS
+90	95	Cdc42	protein	We envisage a complex interplay of equilibria between free and bound, active and inactive Cdc42, TOCA family, and WASP family proteins, facilitating a tightly spatially and temporally regulated pathway requiring numerous simultaneous events in order to achieve appropriate and robust activation of the downstream pathway.	DISCUSS
+97	108	TOCA family	protein_type	We envisage a complex interplay of equilibria between free and bound, active and inactive Cdc42, TOCA family, and WASP family proteins, facilitating a tightly spatially and temporally regulated pathway requiring numerous simultaneous events in order to achieve appropriate and robust activation of the downstream pathway.	DISCUSS
+114	118	WASP	protein_type	We envisage a complex interplay of equilibria between free and bound, active and inactive Cdc42, TOCA family, and WASP family proteins, facilitating a tightly spatially and temporally regulated pathway requiring numerous simultaneous events in order to achieve appropriate and robust activation of the downstream pathway.	DISCUSS
+204	208	WASP	protein_type	Our data are therefore easily reconciled with the dynamic instability models described in relation to the formation of endocytic vesicles and with the current data pertaining to the complex activation of WASP/N-WASP pathways by allosteric and oligomeric effects.	DISCUSS
+209	215	N-WASP	protein	Our data are therefore easily reconciled with the dynamic instability models described in relation to the formation of endocytic vesicles and with the current data pertaining to the complex activation of WASP/N-WASP pathways by allosteric and oligomeric effects.	DISCUSS
+46	51	TOCA1	protein	It is clear from the data presented here that TOCA1 and N-WASP do not bind Cdc42 simultaneously and that N-WASP is likely to outcompete TOCA1 for Cdc42 binding.	DISCUSS
+56	62	N-WASP	protein	It is clear from the data presented here that TOCA1 and N-WASP do not bind Cdc42 simultaneously and that N-WASP is likely to outcompete TOCA1 for Cdc42 binding.	DISCUSS
+75	80	Cdc42	protein	It is clear from the data presented here that TOCA1 and N-WASP do not bind Cdc42 simultaneously and that N-WASP is likely to outcompete TOCA1 for Cdc42 binding.	DISCUSS
+105	111	N-WASP	protein	It is clear from the data presented here that TOCA1 and N-WASP do not bind Cdc42 simultaneously and that N-WASP is likely to outcompete TOCA1 for Cdc42 binding.	DISCUSS
+136	141	TOCA1	protein	It is clear from the data presented here that TOCA1 and N-WASP do not bind Cdc42 simultaneously and that N-WASP is likely to outcompete TOCA1 for Cdc42 binding.	DISCUSS
+146	151	Cdc42	protein	It is clear from the data presented here that TOCA1 and N-WASP do not bind Cdc42 simultaneously and that N-WASP is likely to outcompete TOCA1 for Cdc42 binding.	DISCUSS
+92	96	WASP	protein	We therefore postulate an effector handover mechanism based on current evidence surrounding WASP/N-WASP activation and our model of the Cdc42·HR1TOCA1 complex.	DISCUSS
+97	103	N-WASP	protein	We therefore postulate an effector handover mechanism based on current evidence surrounding WASP/N-WASP activation and our model of the Cdc42·HR1TOCA1 complex.	DISCUSS
+136	150	Cdc42·HR1TOCA1	complex_assembly	We therefore postulate an effector handover mechanism based on current evidence surrounding WASP/N-WASP activation and our model of the Cdc42·HR1TOCA1 complex.	DISCUSS
+24	29	TOCA1	protein	The displacement of the TOCA1 HR1 domain from Cdc42 by N-WASP may represent a unidirectional step in the pathway of Cdc42·N-WASP·TOCA1-dependent actin assembly.	DISCUSS
+30	33	HR1	structure_element	The displacement of the TOCA1 HR1 domain from Cdc42 by N-WASP may represent a unidirectional step in the pathway of Cdc42·N-WASP·TOCA1-dependent actin assembly.	DISCUSS
+46	51	Cdc42	protein	The displacement of the TOCA1 HR1 domain from Cdc42 by N-WASP may represent a unidirectional step in the pathway of Cdc42·N-WASP·TOCA1-dependent actin assembly.	DISCUSS
+55	61	N-WASP	protein	The displacement of the TOCA1 HR1 domain from Cdc42 by N-WASP may represent a unidirectional step in the pathway of Cdc42·N-WASP·TOCA1-dependent actin assembly.	DISCUSS
+116	134	Cdc42·N-WASP·TOCA1	complex_assembly	The displacement of the TOCA1 HR1 domain from Cdc42 by N-WASP may represent a unidirectional step in the pathway of Cdc42·N-WASP·TOCA1-dependent actin assembly.	DISCUSS