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