|
anno_start anno_end anno_text entity_type sentence section |
|
36 44 RORgamma protein Structural determinant for inducing RORgamma specific inverse agonism triggered by a synthetic benzoxazinone ligand TITLE |
|
95 108 benzoxazinone chemical Structural determinant for inducing RORgamma specific inverse agonism triggered by a synthetic benzoxazinone ligand TITLE |
|
4 28 nuclear hormone receptor protein_type The nuclear hormone receptor RORγ regulates transcriptional genes involved in the production of the pro-inflammatory interleukin IL-17 which has been linked to autoimmune diseases such as rheumatoid arthritis, multiple sclerosis and inflammatory bowel disease. ABSTRACT |
|
29 33 RORγ protein The nuclear hormone receptor RORγ regulates transcriptional genes involved in the production of the pro-inflammatory interleukin IL-17 which has been linked to autoimmune diseases such as rheumatoid arthritis, multiple sclerosis and inflammatory bowel disease. ABSTRACT |
|
117 128 interleukin protein_type The nuclear hormone receptor RORγ regulates transcriptional genes involved in the production of the pro-inflammatory interleukin IL-17 which has been linked to autoimmune diseases such as rheumatoid arthritis, multiple sclerosis and inflammatory bowel disease. ABSTRACT |
|
129 134 IL-17 protein_type The nuclear hormone receptor RORγ regulates transcriptional genes involved in the production of the pro-inflammatory interleukin IL-17 which has been linked to autoimmune diseases such as rheumatoid arthritis, multiple sclerosis and inflammatory bowel disease. ABSTRACT |
|
33 37 RORγ protein This transcriptional activity of RORγ is modulated through a protein-protein interaction involving the activation function 2 (AF2) helix on the ligand binding domain of RORγ and a conserved LXXLL helix motif on coactivator proteins. ABSTRACT |
|
103 136 activation function 2 (AF2) helix structure_element This transcriptional activity of RORγ is modulated through a protein-protein interaction involving the activation function 2 (AF2) helix on the ligand binding domain of RORγ and a conserved LXXLL helix motif on coactivator proteins. ABSTRACT |
|
144 165 ligand binding domain structure_element This transcriptional activity of RORγ is modulated through a protein-protein interaction involving the activation function 2 (AF2) helix on the ligand binding domain of RORγ and a conserved LXXLL helix motif on coactivator proteins. ABSTRACT |
|
169 173 RORγ protein This transcriptional activity of RORγ is modulated through a protein-protein interaction involving the activation function 2 (AF2) helix on the ligand binding domain of RORγ and a conserved LXXLL helix motif on coactivator proteins. ABSTRACT |
|
180 189 conserved protein_state This transcriptional activity of RORγ is modulated through a protein-protein interaction involving the activation function 2 (AF2) helix on the ligand binding domain of RORγ and a conserved LXXLL helix motif on coactivator proteins. ABSTRACT |
|
190 207 LXXLL helix motif structure_element This transcriptional activity of RORγ is modulated through a protein-protein interaction involving the activation function 2 (AF2) helix on the ligand binding domain of RORγ and a conserved LXXLL helix motif on coactivator proteins. ABSTRACT |
|
26 30 RORγ protein Our goal was to develop a RORγ specific inverse agonist that would help down regulate pro-inflammatory gene transcription by disrupting the protein protein interaction with coactivator proteins as a therapeutic agent. ABSTRACT |
|
40 55 inverse agonist protein_state Our goal was to develop a RORγ specific inverse agonist that would help down regulate pro-inflammatory gene transcription by disrupting the protein protein interaction with coactivator proteins as a therapeutic agent. ABSTRACT |
|
42 55 benzoxazinone chemical We identified a novel series of synthetic benzoxazinone ligands having an agonist (BIO592) and inverse agonist (BIO399) mode of action in a FRET based assay. ABSTRACT |
|
74 81 agonist protein_state We identified a novel series of synthetic benzoxazinone ligands having an agonist (BIO592) and inverse agonist (BIO399) mode of action in a FRET based assay. ABSTRACT |
|
83 89 BIO592 chemical We identified a novel series of synthetic benzoxazinone ligands having an agonist (BIO592) and inverse agonist (BIO399) mode of action in a FRET based assay. ABSTRACT |
|
95 110 inverse agonist protein_state We identified a novel series of synthetic benzoxazinone ligands having an agonist (BIO592) and inverse agonist (BIO399) mode of action in a FRET based assay. ABSTRACT |
|
112 118 BIO399 chemical We identified a novel series of synthetic benzoxazinone ligands having an agonist (BIO592) and inverse agonist (BIO399) mode of action in a FRET based assay. ABSTRACT |
|
140 156 FRET based assay experimental_method We identified a novel series of synthetic benzoxazinone ligands having an agonist (BIO592) and inverse agonist (BIO399) mode of action in a FRET based assay. ABSTRACT |
|
17 26 AF2 helix structure_element We show that the AF2 helix of RORγ is proteolytically sensitive when inverse agonist BIO399 binds. ABSTRACT |
|
30 34 RORγ protein We show that the AF2 helix of RORγ is proteolytically sensitive when inverse agonist BIO399 binds. ABSTRACT |
|
38 63 proteolytically sensitive protein_state We show that the AF2 helix of RORγ is proteolytically sensitive when inverse agonist BIO399 binds. ABSTRACT |
|
69 84 inverse agonist protein_state We show that the AF2 helix of RORγ is proteolytically sensitive when inverse agonist BIO399 binds. ABSTRACT |
|
85 91 BIO399 chemical We show that the AF2 helix of RORγ is proteolytically sensitive when inverse agonist BIO399 binds. ABSTRACT |
|
6 27 x-ray crystallography experimental_method Using x-ray crystallography we show how small modifications on the benzoxazinone agonist BIO592 trigger inverse agonism of RORγ. ABSTRACT |
|
67 80 benzoxazinone chemical Using x-ray crystallography we show how small modifications on the benzoxazinone agonist BIO592 trigger inverse agonism of RORγ. ABSTRACT |
|
81 88 agonist protein_state Using x-ray crystallography we show how small modifications on the benzoxazinone agonist BIO592 trigger inverse agonism of RORγ. ABSTRACT |
|
89 95 BIO592 chemical Using x-ray crystallography we show how small modifications on the benzoxazinone agonist BIO592 trigger inverse agonism of RORγ. ABSTRACT |
|
123 127 RORγ protein Using x-ray crystallography we show how small modifications on the benzoxazinone agonist BIO592 trigger inverse agonism of RORγ. ABSTRACT |
|
9 31 in vivo reporter assay experimental_method Using an in vivo reporter assay, we show that the inverse agonist BIO399 displayed specificity for RORγ over ROR sub-family members α and β. ABSTRACT |
|
50 65 inverse agonist protein_state Using an in vivo reporter assay, we show that the inverse agonist BIO399 displayed specificity for RORγ over ROR sub-family members α and β. ABSTRACT |
|
66 72 BIO399 chemical Using an in vivo reporter assay, we show that the inverse agonist BIO399 displayed specificity for RORγ over ROR sub-family members α and β. ABSTRACT |
|
99 103 RORγ protein Using an in vivo reporter assay, we show that the inverse agonist BIO399 displayed specificity for RORγ over ROR sub-family members α and β. ABSTRACT |
|
109 112 ROR protein_type Using an in vivo reporter assay, we show that the inverse agonist BIO399 displayed specificity for RORγ over ROR sub-family members α and β. ABSTRACT |
|
132 133 α protein Using an in vivo reporter assay, we show that the inverse agonist BIO399 displayed specificity for RORγ over ROR sub-family members α and β. ABSTRACT |
|
138 139 β protein Using an in vivo reporter assay, we show that the inverse agonist BIO399 displayed specificity for RORγ over ROR sub-family members α and β. ABSTRACT |
|
14 27 benzoxazinone chemical The synthetic benzoxazinone ligands identified in our FRET assay have an agonist (BIO592) or inverse agonist (BIO399) effect by stabilizing or destabilizing the agonist conformation of RORγ. ABSTRACT |
|
54 64 FRET assay experimental_method The synthetic benzoxazinone ligands identified in our FRET assay have an agonist (BIO592) or inverse agonist (BIO399) effect by stabilizing or destabilizing the agonist conformation of RORγ. ABSTRACT |
|
73 80 agonist protein_state The synthetic benzoxazinone ligands identified in our FRET assay have an agonist (BIO592) or inverse agonist (BIO399) effect by stabilizing or destabilizing the agonist conformation of RORγ. ABSTRACT |
|
82 88 BIO592 chemical The synthetic benzoxazinone ligands identified in our FRET assay have an agonist (BIO592) or inverse agonist (BIO399) effect by stabilizing or destabilizing the agonist conformation of RORγ. ABSTRACT |
|
93 108 inverse agonist protein_state The synthetic benzoxazinone ligands identified in our FRET assay have an agonist (BIO592) or inverse agonist (BIO399) effect by stabilizing or destabilizing the agonist conformation of RORγ. ABSTRACT |
|
110 116 BIO399 chemical The synthetic benzoxazinone ligands identified in our FRET assay have an agonist (BIO592) or inverse agonist (BIO399) effect by stabilizing or destabilizing the agonist conformation of RORγ. ABSTRACT |
|
161 168 agonist protein_state The synthetic benzoxazinone ligands identified in our FRET assay have an agonist (BIO592) or inverse agonist (BIO399) effect by stabilizing or destabilizing the agonist conformation of RORγ. ABSTRACT |
|
185 189 RORγ protein The synthetic benzoxazinone ligands identified in our FRET assay have an agonist (BIO592) or inverse agonist (BIO399) effect by stabilizing or destabilizing the agonist conformation of RORγ. ABSTRACT |
|
35 44 AF2 helix structure_element The proteolytic sensitivity of the AF2 helix of RORγ demonstrates that it destabilizes upon BIO399 inverse agonist binding perturbing the coactivator protein binding site. ABSTRACT |
|
48 52 RORγ protein The proteolytic sensitivity of the AF2 helix of RORγ demonstrates that it destabilizes upon BIO399 inverse agonist binding perturbing the coactivator protein binding site. ABSTRACT |
|
92 98 BIO399 chemical The proteolytic sensitivity of the AF2 helix of RORγ demonstrates that it destabilizes upon BIO399 inverse agonist binding perturbing the coactivator protein binding site. ABSTRACT |
|
99 114 inverse agonist protein_state The proteolytic sensitivity of the AF2 helix of RORγ demonstrates that it destabilizes upon BIO399 inverse agonist binding perturbing the coactivator protein binding site. ABSTRACT |
|
138 170 coactivator protein binding site site The proteolytic sensitivity of the AF2 helix of RORγ demonstrates that it destabilizes upon BIO399 inverse agonist binding perturbing the coactivator protein binding site. ABSTRACT |
|
4 28 structural investigation experimental_method Our structural investigation of the BIO592 agonist and BIO399 inverse agonist structures identified residue Met358 on RORγ as the trigger for RORγ specific inverse agonism. ABSTRACT |
|
36 42 BIO592 chemical Our structural investigation of the BIO592 agonist and BIO399 inverse agonist structures identified residue Met358 on RORγ as the trigger for RORγ specific inverse agonism. ABSTRACT |
|
43 50 agonist protein_state Our structural investigation of the BIO592 agonist and BIO399 inverse agonist structures identified residue Met358 on RORγ as the trigger for RORγ specific inverse agonism. ABSTRACT |
|
55 61 BIO399 chemical Our structural investigation of the BIO592 agonist and BIO399 inverse agonist structures identified residue Met358 on RORγ as the trigger for RORγ specific inverse agonism. ABSTRACT |
|
62 77 inverse agonist protein_state Our structural investigation of the BIO592 agonist and BIO399 inverse agonist structures identified residue Met358 on RORγ as the trigger for RORγ specific inverse agonism. ABSTRACT |
|
78 88 structures evidence Our structural investigation of the BIO592 agonist and BIO399 inverse agonist structures identified residue Met358 on RORγ as the trigger for RORγ specific inverse agonism. ABSTRACT |
|
108 114 Met358 residue_name_number Our structural investigation of the BIO592 agonist and BIO399 inverse agonist structures identified residue Met358 on RORγ as the trigger for RORγ specific inverse agonism. ABSTRACT |
|
118 122 RORγ protein Our structural investigation of the BIO592 agonist and BIO399 inverse agonist structures identified residue Met358 on RORγ as the trigger for RORγ specific inverse agonism. ABSTRACT |
|
142 146 RORγ protein Our structural investigation of the BIO592 agonist and BIO399 inverse agonist structures identified residue Met358 on RORγ as the trigger for RORγ specific inverse agonism. ABSTRACT |
|
0 38 Retinoid-related orphan receptor gamma protein Retinoid-related orphan receptor gamma (RORγ) is a transcription factor belonging to a sub-family of nuclear receptors that includes two closely related members RORα and RORβ. INTRO |
|
40 44 RORγ protein Retinoid-related orphan receptor gamma (RORγ) is a transcription factor belonging to a sub-family of nuclear receptors that includes two closely related members RORα and RORβ. INTRO |
|
51 71 transcription factor protein_type Retinoid-related orphan receptor gamma (RORγ) is a transcription factor belonging to a sub-family of nuclear receptors that includes two closely related members RORα and RORβ. INTRO |
|
101 118 nuclear receptors protein_type Retinoid-related orphan receptor gamma (RORγ) is a transcription factor belonging to a sub-family of nuclear receptors that includes two closely related members RORα and RORβ. INTRO |
|
161 165 RORα protein Retinoid-related orphan receptor gamma (RORγ) is a transcription factor belonging to a sub-family of nuclear receptors that includes two closely related members RORα and RORβ. INTRO |
|
170 174 RORβ protein Retinoid-related orphan receptor gamma (RORγ) is a transcription factor belonging to a sub-family of nuclear receptors that includes two closely related members RORα and RORβ. INTRO |
|
68 72 RORs protein_type Even though a high degree of sequence similarity exists between the RORs, their functional roles in regulation for physiological processes involved in development and immunity are distinct. INTRO |
|
20 24 RORγ protein During development, RORγ regulates the transcriptional genes involved in the functioning of multiple pro-inflammatory lymphocyte lineages including T helper cells (TH17cells) which are necessary for IL-17 production. INTRO |
|
199 204 IL-17 protein_type During development, RORγ regulates the transcriptional genes involved in the functioning of multiple pro-inflammatory lymphocyte lineages including T helper cells (TH17cells) which are necessary for IL-17 production. INTRO |
|
0 5 IL-17 protein_type IL-17 is a pro-inflammatory interleukin linked to autoimmune diseases such as rheumatoid arthritis, multiple sclerosis and inflammatory bowel disease; making its transcriptional regulation through RORγ an attractive therapeutic target. INTRO |
|
28 39 interleukin protein_type IL-17 is a pro-inflammatory interleukin linked to autoimmune diseases such as rheumatoid arthritis, multiple sclerosis and inflammatory bowel disease; making its transcriptional regulation through RORγ an attractive therapeutic target. INTRO |
|
197 201 RORγ protein IL-17 is a pro-inflammatory interleukin linked to autoimmune diseases such as rheumatoid arthritis, multiple sclerosis and inflammatory bowel disease; making its transcriptional regulation through RORγ an attractive therapeutic target. INTRO |
|
0 4 RORγ protein RORγ consists of an N-terminal DNA binding domain (DBD) connected to a C-terminal ligand binding domain (LBD) via a flexible hinge region. INTRO |
|
31 49 DNA binding domain structure_element RORγ consists of an N-terminal DNA binding domain (DBD) connected to a C-terminal ligand binding domain (LBD) via a flexible hinge region. INTRO |
|
51 54 DBD structure_element RORγ consists of an N-terminal DNA binding domain (DBD) connected to a C-terminal ligand binding domain (LBD) via a flexible hinge region. INTRO |
|
82 103 ligand binding domain structure_element RORγ consists of an N-terminal DNA binding domain (DBD) connected to a C-terminal ligand binding domain (LBD) via a flexible hinge region. INTRO |
|
105 108 LBD structure_element RORγ consists of an N-terminal DNA binding domain (DBD) connected to a C-terminal ligand binding domain (LBD) via a flexible hinge region. INTRO |
|
125 137 hinge region structure_element RORγ consists of an N-terminal DNA binding domain (DBD) connected to a C-terminal ligand binding domain (LBD) via a flexible hinge region. INTRO |
|
4 7 DBD structure_element The DBD is composed of two zinc fingers that allow it to interact with specifically encoded regions on the DNA called the nuclear receptor response elements. INTRO |
|
27 39 zinc fingers structure_element The DBD is composed of two zinc fingers that allow it to interact with specifically encoded regions on the DNA called the nuclear receptor response elements. INTRO |
|
122 156 nuclear receptor response elements structure_element The DBD is composed of two zinc fingers that allow it to interact with specifically encoded regions on the DNA called the nuclear receptor response elements. INTRO |
|
4 7 LBD structure_element The LBD consists of a coactivator protein binding pocket and a hydrophobic ligand binding site (LBS) which are responsible for regulating transcription. INTRO |
|
22 56 coactivator protein binding pocket site The LBD consists of a coactivator protein binding pocket and a hydrophobic ligand binding site (LBS) which are responsible for regulating transcription. INTRO |
|
63 94 hydrophobic ligand binding site site The LBD consists of a coactivator protein binding pocket and a hydrophobic ligand binding site (LBS) which are responsible for regulating transcription. INTRO |
|
96 99 LBS site The LBD consists of a coactivator protein binding pocket and a hydrophobic ligand binding site (LBS) which are responsible for regulating transcription. INTRO |
|
4 30 coactivator binding pocket site The coactivator binding pocket of RORγ recognizes a conserved helix motif LXXLL (where X can be any amino acid) on transcriptional coactivator complexes and recruits it to activate transcription. INTRO |
|
34 38 RORγ protein The coactivator binding pocket of RORγ recognizes a conserved helix motif LXXLL (where X can be any amino acid) on transcriptional coactivator complexes and recruits it to activate transcription. INTRO |
|
52 61 conserved protein_state The coactivator binding pocket of RORγ recognizes a conserved helix motif LXXLL (where X can be any amino acid) on transcriptional coactivator complexes and recruits it to activate transcription. INTRO |
|
62 79 helix motif LXXLL structure_element The coactivator binding pocket of RORγ recognizes a conserved helix motif LXXLL (where X can be any amino acid) on transcriptional coactivator complexes and recruits it to activate transcription. INTRO |
|
11 36 nuclear hormone receptors protein_type Like other nuclear hormone receptors, RORγ’s helix12 which makes up the C-termini of the LBD is an essential part of the coactivator binding pocket and is commonly referred to as the activation function helix 2 (AF2). INTRO |
|
38 42 RORγ protein Like other nuclear hormone receptors, RORγ’s helix12 which makes up the C-termini of the LBD is an essential part of the coactivator binding pocket and is commonly referred to as the activation function helix 2 (AF2). INTRO |
|
45 52 helix12 structure_element Like other nuclear hormone receptors, RORγ’s helix12 which makes up the C-termini of the LBD is an essential part of the coactivator binding pocket and is commonly referred to as the activation function helix 2 (AF2). INTRO |
|
89 92 LBD structure_element Like other nuclear hormone receptors, RORγ’s helix12 which makes up the C-termini of the LBD is an essential part of the coactivator binding pocket and is commonly referred to as the activation function helix 2 (AF2). INTRO |
|
121 147 coactivator binding pocket site Like other nuclear hormone receptors, RORγ’s helix12 which makes up the C-termini of the LBD is an essential part of the coactivator binding pocket and is commonly referred to as the activation function helix 2 (AF2). INTRO |
|
183 210 activation function helix 2 structure_element Like other nuclear hormone receptors, RORγ’s helix12 which makes up the C-termini of the LBD is an essential part of the coactivator binding pocket and is commonly referred to as the activation function helix 2 (AF2). INTRO |
|
212 215 AF2 structure_element Like other nuclear hormone receptors, RORγ’s helix12 which makes up the C-termini of the LBD is an essential part of the coactivator binding pocket and is commonly referred to as the activation function helix 2 (AF2). INTRO |
|
3 7 RORγ protein In RORγ, the conformation of the AF2 helix required to form the coactivator binding pocket is mediated by a salt bridge between His479 and Tyr502 in addition to π- π interactions between Tyr502 and Phe506. INTRO |
|
33 42 AF2 helix structure_element In RORγ, the conformation of the AF2 helix required to form the coactivator binding pocket is mediated by a salt bridge between His479 and Tyr502 in addition to π- π interactions between Tyr502 and Phe506. INTRO |
|
64 90 coactivator binding pocket site In RORγ, the conformation of the AF2 helix required to form the coactivator binding pocket is mediated by a salt bridge between His479 and Tyr502 in addition to π- π interactions between Tyr502 and Phe506. INTRO |
|
108 119 salt bridge bond_interaction In RORγ, the conformation of the AF2 helix required to form the coactivator binding pocket is mediated by a salt bridge between His479 and Tyr502 in addition to π- π interactions between Tyr502 and Phe506. INTRO |
|
128 134 His479 residue_name_number In RORγ, the conformation of the AF2 helix required to form the coactivator binding pocket is mediated by a salt bridge between His479 and Tyr502 in addition to π- π interactions between Tyr502 and Phe506. INTRO |
|
139 145 Tyr502 residue_name_number In RORγ, the conformation of the AF2 helix required to form the coactivator binding pocket is mediated by a salt bridge between His479 and Tyr502 in addition to π- π interactions between Tyr502 and Phe506. INTRO |
|
161 178 π- π interactions bond_interaction In RORγ, the conformation of the AF2 helix required to form the coactivator binding pocket is mediated by a salt bridge between His479 and Tyr502 in addition to π- π interactions between Tyr502 and Phe506. INTRO |
|
187 193 Tyr502 residue_name_number In RORγ, the conformation of the AF2 helix required to form the coactivator binding pocket is mediated by a salt bridge between His479 and Tyr502 in addition to π- π interactions between Tyr502 and Phe506. INTRO |
|
198 204 Phe506 residue_name_number In RORγ, the conformation of the AF2 helix required to form the coactivator binding pocket is mediated by a salt bridge between His479 and Tyr502 in addition to π- π interactions between Tyr502 and Phe506. INTRO |
|
24 33 AF2 helix structure_element The conformation of the AF2 helix can be modulated through targeted ligands which bind the LBS and increase the binding of the coactivator protein (agonists) or disrupt binding (inverse agonists) thereby enhancing or inhibiting transcription. INTRO |
|
91 94 LBS site The conformation of the AF2 helix can be modulated through targeted ligands which bind the LBS and increase the binding of the coactivator protein (agonists) or disrupt binding (inverse agonists) thereby enhancing or inhibiting transcription. INTRO |
|
6 10 RORγ protein Since RORγ has been demonstrated to play an important role in pro-inflammatory gene expression patterns implicated in several major autoimmune diseases, our aim was to develop RORγ inverse agonists that would help down regulate pro-inflammatory gene transcription. INTRO |
|
176 180 RORγ protein Since RORγ has been demonstrated to play an important role in pro-inflammatory gene expression patterns implicated in several major autoimmune diseases, our aim was to develop RORγ inverse agonists that would help down regulate pro-inflammatory gene transcription. INTRO |
|
0 12 FRET results evidence FRET results for agonist BIO592 (a) and Inverse Agonist BIO399 (b) FIG |
|
17 24 agonist protein_state FRET results for agonist BIO592 (a) and Inverse Agonist BIO399 (b) FIG |
|
25 31 BIO592 chemical FRET results for agonist BIO592 (a) and Inverse Agonist BIO399 (b) FIG |
|
40 55 Inverse Agonist protein_state FRET results for agonist BIO592 (a) and Inverse Agonist BIO399 (b) FIG |
|
56 62 BIO399 chemical FRET results for agonist BIO592 (a) and Inverse Agonist BIO399 (b) FIG |
|
52 65 benzoxazinone chemical Here we present the identification of two synthetic benzoxazinone RORγ ligands, a weak agonist BIO592 (Fig. 1a) and an inverse agonist BIO399 (Fig. 1b) which were identified using a Fluorescence Resonance Energy transfer (FRET) based assay that monitored coactivator peptide recruitment. INTRO |
|
66 70 RORγ protein Here we present the identification of two synthetic benzoxazinone RORγ ligands, a weak agonist BIO592 (Fig. 1a) and an inverse agonist BIO399 (Fig. 1b) which were identified using a Fluorescence Resonance Energy transfer (FRET) based assay that monitored coactivator peptide recruitment. INTRO |
|
87 94 agonist protein_state Here we present the identification of two synthetic benzoxazinone RORγ ligands, a weak agonist BIO592 (Fig. 1a) and an inverse agonist BIO399 (Fig. 1b) which were identified using a Fluorescence Resonance Energy transfer (FRET) based assay that monitored coactivator peptide recruitment. INTRO |
|
95 101 BIO592 chemical Here we present the identification of two synthetic benzoxazinone RORγ ligands, a weak agonist BIO592 (Fig. 1a) and an inverse agonist BIO399 (Fig. 1b) which were identified using a Fluorescence Resonance Energy transfer (FRET) based assay that monitored coactivator peptide recruitment. INTRO |
|
119 134 inverse agonist protein_state Here we present the identification of two synthetic benzoxazinone RORγ ligands, a weak agonist BIO592 (Fig. 1a) and an inverse agonist BIO399 (Fig. 1b) which were identified using a Fluorescence Resonance Energy transfer (FRET) based assay that monitored coactivator peptide recruitment. INTRO |
|
135 141 BIO399 chemical Here we present the identification of two synthetic benzoxazinone RORγ ligands, a weak agonist BIO592 (Fig. 1a) and an inverse agonist BIO399 (Fig. 1b) which were identified using a Fluorescence Resonance Energy transfer (FRET) based assay that monitored coactivator peptide recruitment. INTRO |
|
182 239 Fluorescence Resonance Energy transfer (FRET) based assay experimental_method Here we present the identification of two synthetic benzoxazinone RORγ ligands, a weak agonist BIO592 (Fig. 1a) and an inverse agonist BIO399 (Fig. 1b) which were identified using a Fluorescence Resonance Energy transfer (FRET) based assay that monitored coactivator peptide recruitment. INTRO |
|
6 25 partial proteolysis experimental_method Using partial proteolysis in combination with mass spectrometry analysis we demonstrate that the AF2 helix of RORγ destabilizes upon BIO399 (inverse agonist) binding. INTRO |
|
46 63 mass spectrometry experimental_method Using partial proteolysis in combination with mass spectrometry analysis we demonstrate that the AF2 helix of RORγ destabilizes upon BIO399 (inverse agonist) binding. INTRO |
|
97 106 AF2 helix structure_element Using partial proteolysis in combination with mass spectrometry analysis we demonstrate that the AF2 helix of RORγ destabilizes upon BIO399 (inverse agonist) binding. INTRO |
|
110 114 RORγ protein Using partial proteolysis in combination with mass spectrometry analysis we demonstrate that the AF2 helix of RORγ destabilizes upon BIO399 (inverse agonist) binding. INTRO |
|
133 139 BIO399 chemical Using partial proteolysis in combination with mass spectrometry analysis we demonstrate that the AF2 helix of RORγ destabilizes upon BIO399 (inverse agonist) binding. INTRO |
|
141 156 inverse agonist protein_state Using partial proteolysis in combination with mass spectrometry analysis we demonstrate that the AF2 helix of RORγ destabilizes upon BIO399 (inverse agonist) binding. INTRO |
|
19 32 binding modes evidence Finally, comparing binding modes of our benzoxazinone RORγ crystal structures to other ROR structures, we hypothesize a new mode of action for achieving inverse agonism and selectivity. INTRO |
|
40 53 benzoxazinone chemical Finally, comparing binding modes of our benzoxazinone RORγ crystal structures to other ROR structures, we hypothesize a new mode of action for achieving inverse agonism and selectivity. INTRO |
|
54 58 RORγ protein Finally, comparing binding modes of our benzoxazinone RORγ crystal structures to other ROR structures, we hypothesize a new mode of action for achieving inverse agonism and selectivity. INTRO |
|
59 77 crystal structures evidence Finally, comparing binding modes of our benzoxazinone RORγ crystal structures to other ROR structures, we hypothesize a new mode of action for achieving inverse agonism and selectivity. INTRO |
|
87 90 ROR protein_type Finally, comparing binding modes of our benzoxazinone RORγ crystal structures to other ROR structures, we hypothesize a new mode of action for achieving inverse agonism and selectivity. INTRO |
|
91 101 structures evidence Finally, comparing binding modes of our benzoxazinone RORγ crystal structures to other ROR structures, we hypothesize a new mode of action for achieving inverse agonism and selectivity. INTRO |
|
8 24 FRET based assay experimental_method Using a FRET based assay we discovered agonist BIO592 (Fig. 1a) which increased the coactivator peptide TRAP220 recruitment to RORγ (EC50 0f 58nM and Emax of 130 %) and a potent inverse agonist BIO399 (Fig. 1b) which inhibited coactivator recruitment (IC50: 4.7nM). RESULTS |
|
39 46 agonist protein_state Using a FRET based assay we discovered agonist BIO592 (Fig. 1a) which increased the coactivator peptide TRAP220 recruitment to RORγ (EC50 0f 58nM and Emax of 130 %) and a potent inverse agonist BIO399 (Fig. 1b) which inhibited coactivator recruitment (IC50: 4.7nM). RESULTS |
|
47 53 BIO592 chemical Using a FRET based assay we discovered agonist BIO592 (Fig. 1a) which increased the coactivator peptide TRAP220 recruitment to RORγ (EC50 0f 58nM and Emax of 130 %) and a potent inverse agonist BIO399 (Fig. 1b) which inhibited coactivator recruitment (IC50: 4.7nM). RESULTS |
|
104 111 TRAP220 chemical Using a FRET based assay we discovered agonist BIO592 (Fig. 1a) which increased the coactivator peptide TRAP220 recruitment to RORγ (EC50 0f 58nM and Emax of 130 %) and a potent inverse agonist BIO399 (Fig. 1b) which inhibited coactivator recruitment (IC50: 4.7nM). RESULTS |
|
127 131 RORγ protein Using a FRET based assay we discovered agonist BIO592 (Fig. 1a) which increased the coactivator peptide TRAP220 recruitment to RORγ (EC50 0f 58nM and Emax of 130 %) and a potent inverse agonist BIO399 (Fig. 1b) which inhibited coactivator recruitment (IC50: 4.7nM). RESULTS |
|
133 137 EC50 evidence Using a FRET based assay we discovered agonist BIO592 (Fig. 1a) which increased the coactivator peptide TRAP220 recruitment to RORγ (EC50 0f 58nM and Emax of 130 %) and a potent inverse agonist BIO399 (Fig. 1b) which inhibited coactivator recruitment (IC50: 4.7nM). RESULTS |
|
150 154 Emax evidence Using a FRET based assay we discovered agonist BIO592 (Fig. 1a) which increased the coactivator peptide TRAP220 recruitment to RORγ (EC50 0f 58nM and Emax of 130 %) and a potent inverse agonist BIO399 (Fig. 1b) which inhibited coactivator recruitment (IC50: 4.7nM). RESULTS |
|
178 193 inverse agonist protein_state Using a FRET based assay we discovered agonist BIO592 (Fig. 1a) which increased the coactivator peptide TRAP220 recruitment to RORγ (EC50 0f 58nM and Emax of 130 %) and a potent inverse agonist BIO399 (Fig. 1b) which inhibited coactivator recruitment (IC50: 4.7nM). RESULTS |
|
194 200 BIO399 chemical Using a FRET based assay we discovered agonist BIO592 (Fig. 1a) which increased the coactivator peptide TRAP220 recruitment to RORγ (EC50 0f 58nM and Emax of 130 %) and a potent inverse agonist BIO399 (Fig. 1b) which inhibited coactivator recruitment (IC50: 4.7nM). RESULTS |
|
252 256 IC50 evidence Using a FRET based assay we discovered agonist BIO592 (Fig. 1a) which increased the coactivator peptide TRAP220 recruitment to RORγ (EC50 0f 58nM and Emax of 130 %) and a potent inverse agonist BIO399 (Fig. 1b) which inhibited coactivator recruitment (IC50: 4.7nM). RESULTS |
|
53 60 agonist protein_state Interestingly, the structural difference between the agonist BIO592 and inverse agonist BIO399 was minor |
|
61 67 BIO592 chemical Interestingly, the structural difference between the agonist BIO592 and inverse agonist BIO399 was minor |
|
72 87 inverse agonist protein_state Interestingly, the structural difference between the agonist BIO592 and inverse agonist BIO399 was minor |
|
88 94 BIO399 chemical Interestingly, the structural difference between the agonist BIO592 and inverse agonist BIO399 was minor |
|
115 150 2,3-dihydrobenzo[1,4]oxazepin-4-one chemical Interestingly, the structural difference between the agonist BIO592 and inverse agonist BIO399 was minor |
|
166 172 BIO399 chemical Interestingly, the structural difference between the agonist BIO592 and inverse agonist BIO399 was minor |
|
203 226 benzo[1,4]oxazine-3-one chemical Interestingly, the structural difference between the agonist BIO592 and inverse agonist BIO399 was minor |
|
242 248 BIO592 chemical Interestingly, the structural difference between the agonist BIO592 and inverse agonist BIO399 was minor |
|
152 158 BIO592 chemical In order to understand how small changes in the core ring system leads to inverse agonism, we wanted to structurally determine the binding mode of both BIO592 and BIO399 in the LBS of RORγ using x-ray crystallography. RESULTS |
|
163 169 BIO399 chemical In order to understand how small changes in the core ring system leads to inverse agonism, we wanted to structurally determine the binding mode of both BIO592 and BIO399 in the LBS of RORγ using x-ray crystallography. RESULTS |
|
177 180 LBS site In order to understand how small changes in the core ring system leads to inverse agonism, we wanted to structurally determine the binding mode of both BIO592 and BIO399 in the LBS of RORγ using x-ray crystallography. RESULTS |
|
184 188 RORγ protein In order to understand how small changes in the core ring system leads to inverse agonism, we wanted to structurally determine the binding mode of both BIO592 and BIO399 in the LBS of RORγ using x-ray crystallography. RESULTS |
|
195 216 x-ray crystallography experimental_method In order to understand how small changes in the core ring system leads to inverse agonism, we wanted to structurally determine the binding mode of both BIO592 and BIO399 in the LBS of RORγ using x-ray crystallography. RESULTS |
|
0 9 Structure evidence Structure of the RORγ518-BIO592-EBI96 ternary complex is in a transcriptionally active conformation RESULTS |
|
17 37 RORγ518-BIO592-EBI96 complex_assembly Structure of the RORγ518-BIO592-EBI96 ternary complex is in a transcriptionally active conformation RESULTS |
|
80 86 active protein_state Structure of the RORγ518-BIO592-EBI96 ternary complex is in a transcriptionally active conformation RESULTS |
|
7 24 ternary structure evidence a The ternary structure of RORγ518 BIO592 and EBI96. FIG |
|
28 35 RORγ518 protein a The ternary structure of RORγ518 BIO592 and EBI96. FIG |
|
36 42 BIO592 chemical a The ternary structure of RORγ518 BIO592 and EBI96. FIG |
|
47 52 EBI96 chemical a The ternary structure of RORγ518 BIO592 and EBI96. FIG |
|
2 6 RORγ protein b RORγ AF2 helix in the agonist conformation. FIG |
|
7 16 AF2 helix structure_element b RORγ AF2 helix in the agonist conformation. FIG |
|
24 31 agonist protein_state b RORγ AF2 helix in the agonist conformation. FIG |
|
2 7 EBI96 chemical c EBI96 coactivator peptide bound in the coactivator pocket of RORγ FIG |
|
28 36 bound in protein_state c EBI96 coactivator peptide bound in the coactivator pocket of RORγ FIG |
|
41 59 coactivator pocket site c EBI96 coactivator peptide bound in the coactivator pocket of RORγ FIG |
|
63 67 RORγ protein c EBI96 coactivator peptide bound in the coactivator pocket of RORγ FIG |
|
0 7 RORγ518 protein RORγ518 bound to agonist BIO592 was crystallized with a truncated form of the coactivator peptide EBI96 to a resolution of 2.6 Å (Fig. 2a). RESULTS |
|
8 16 bound to protein_state RORγ518 bound to agonist BIO592 was crystallized with a truncated form of the coactivator peptide EBI96 to a resolution of 2.6 Å (Fig. 2a). RESULTS |
|
17 24 agonist protein_state RORγ518 bound to agonist BIO592 was crystallized with a truncated form of the coactivator peptide EBI96 to a resolution of 2.6 Å (Fig. 2a). RESULTS |
|
25 31 BIO592 chemical RORγ518 bound to agonist BIO592 was crystallized with a truncated form of the coactivator peptide EBI96 to a resolution of 2.6 Å (Fig. 2a). RESULTS |
|
36 48 crystallized experimental_method RORγ518 bound to agonist BIO592 was crystallized with a truncated form of the coactivator peptide EBI96 to a resolution of 2.6 Å (Fig. 2a). RESULTS |
|
56 65 truncated protein_state RORγ518 bound to agonist BIO592 was crystallized with a truncated form of the coactivator peptide EBI96 to a resolution of 2.6 Å (Fig. 2a). RESULTS |
|
98 103 EBI96 chemical RORγ518 bound to agonist BIO592 was crystallized with a truncated form of the coactivator peptide EBI96 to a resolution of 2.6 Å (Fig. 2a). RESULTS |
|
4 13 structure evidence The structure of the ternary complex had features similar to other ROR agonist coactivator structures in a transcriptionally active canonical three layer helix fold with the AF2 helix in the agonist conformation. RESULTS |
|
67 70 ROR protein_type The structure of the ternary complex had features similar to other ROR agonist coactivator structures in a transcriptionally active canonical three layer helix fold with the AF2 helix in the agonist conformation. RESULTS |
|
71 78 agonist protein_state The structure of the ternary complex had features similar to other ROR agonist coactivator structures in a transcriptionally active canonical three layer helix fold with the AF2 helix in the agonist conformation. RESULTS |
|
91 101 structures evidence The structure of the ternary complex had features similar to other ROR agonist coactivator structures in a transcriptionally active canonical three layer helix fold with the AF2 helix in the agonist conformation. RESULTS |
|
107 131 transcriptionally active protein_state The structure of the ternary complex had features similar to other ROR agonist coactivator structures in a transcriptionally active canonical three layer helix fold with the AF2 helix in the agonist conformation. RESULTS |
|
132 164 canonical three layer helix fold protein_state The structure of the ternary complex had features similar to other ROR agonist coactivator structures in a transcriptionally active canonical three layer helix fold with the AF2 helix in the agonist conformation. RESULTS |
|
174 183 AF2 helix structure_element The structure of the ternary complex had features similar to other ROR agonist coactivator structures in a transcriptionally active canonical three layer helix fold with the AF2 helix in the agonist conformation. RESULTS |
|
191 198 agonist protein_state The structure of the ternary complex had features similar to other ROR agonist coactivator structures in a transcriptionally active canonical three layer helix fold with the AF2 helix in the agonist conformation. RESULTS |
|
4 11 agonist protein_state The agonist conformation is stabilized by a hydrogen bond between His479 and Tyr502, in addition to π-π interactions between His479, Tyr502 and Phe506 (Fig. 2b). RESULTS |
|
44 57 hydrogen bond bond_interaction The agonist conformation is stabilized by a hydrogen bond between His479 and Tyr502, in addition to π-π interactions between His479, Tyr502 and Phe506 (Fig. 2b). RESULTS |
|
66 72 His479 residue_name_number The agonist conformation is stabilized by a hydrogen bond between His479 and Tyr502, in addition to π-π interactions between His479, Tyr502 and Phe506 (Fig. 2b). RESULTS |
|
77 83 Tyr502 residue_name_number The agonist conformation is stabilized by a hydrogen bond between His479 and Tyr502, in addition to π-π interactions between His479, Tyr502 and Phe506 (Fig. 2b). RESULTS |
|
100 116 π-π interactions bond_interaction The agonist conformation is stabilized by a hydrogen bond between His479 and Tyr502, in addition to π-π interactions between His479, Tyr502 and Phe506 (Fig. 2b). RESULTS |
|
125 131 His479 residue_name_number The agonist conformation is stabilized by a hydrogen bond between His479 and Tyr502, in addition to π-π interactions between His479, Tyr502 and Phe506 (Fig. 2b). RESULTS |
|
133 139 Tyr502 residue_name_number The agonist conformation is stabilized by a hydrogen bond between His479 and Tyr502, in addition to π-π interactions between His479, Tyr502 and Phe506 (Fig. 2b). RESULTS |
|
144 150 Phe506 residue_name_number The agonist conformation is stabilized by a hydrogen bond between His479 and Tyr502, in addition to π-π interactions between His479, Tyr502 and Phe506 (Fig. 2b). RESULTS |
|
4 17 hydrogen bond bond_interaction The hydrogen bond between His479 and Tyr502 has been reported to be critical for RORγ agonist activity. RESULTS |
|
26 32 His479 residue_name_number The hydrogen bond between His479 and Tyr502 has been reported to be critical for RORγ agonist activity. RESULTS |
|
37 43 Tyr502 residue_name_number The hydrogen bond between His479 and Tyr502 has been reported to be critical for RORγ agonist activity. RESULTS |
|
81 85 RORγ protein The hydrogen bond between His479 and Tyr502 has been reported to be critical for RORγ agonist activity. RESULTS |
|
86 93 agonist protein_state The hydrogen bond between His479 and Tyr502 has been reported to be critical for RORγ agonist activity. RESULTS |
|
36 47 mutagenesis experimental_method Disrupting this interaction through mutagenesis reduced transcriptional activity of RORγ. RESULTS |
|
84 88 RORγ protein Disrupting this interaction through mutagenesis reduced transcriptional activity of RORγ. RESULTS |
|
82 91 AF2 helix structure_element This reduced transcriptional activity has been attributed to the inability of the AF2 helix to complete the formation of the coactivator binding pocket necessary for coactivator proteins to bind. RESULTS |
|
125 151 coactivator binding pocket site This reduced transcriptional activity has been attributed to the inability of the AF2 helix to complete the formation of the coactivator binding pocket necessary for coactivator proteins to bind. RESULTS |
|
0 16 Electron density evidence Electron density for the coactivator peptide EBI96 was observed for residues EFPYLLSLLG which formed a α-helix stabilized through hydrophobic interactions with the coactivator binding pocket on RORγ (Fig. 2c). RESULTS |
|
45 50 EBI96 chemical Electron density for the coactivator peptide EBI96 was observed for residues EFPYLLSLLG which formed a α-helix stabilized through hydrophobic interactions with the coactivator binding pocket on RORγ (Fig. 2c). RESULTS |
|
77 87 EFPYLLSLLG structure_element Electron density for the coactivator peptide EBI96 was observed for residues EFPYLLSLLG which formed a α-helix stabilized through hydrophobic interactions with the coactivator binding pocket on RORγ (Fig. 2c). RESULTS |
|
103 110 α-helix structure_element Electron density for the coactivator peptide EBI96 was observed for residues EFPYLLSLLG which formed a α-helix stabilized through hydrophobic interactions with the coactivator binding pocket on RORγ (Fig. 2c). RESULTS |
|
130 154 hydrophobic interactions bond_interaction Electron density for the coactivator peptide EBI96 was observed for residues EFPYLLSLLG which formed a α-helix stabilized through hydrophobic interactions with the coactivator binding pocket on RORγ (Fig. 2c). RESULTS |
|
164 190 coactivator binding pocket site Electron density for the coactivator peptide EBI96 was observed for residues EFPYLLSLLG which formed a α-helix stabilized through hydrophobic interactions with the coactivator binding pocket on RORγ (Fig. 2c). RESULTS |
|
194 198 RORγ protein Electron density for the coactivator peptide EBI96 was observed for residues EFPYLLSLLG which formed a α-helix stabilized through hydrophobic interactions with the coactivator binding pocket on RORγ (Fig. 2c). RESULTS |
|
49 58 conserved protein_state This interaction is further stabilized through a conserved charged clamp wherein the backbone amide of Tyr7 and carbonyl of Leu11 of EBI96 form hydrogen bonds with Glu504 (helix12) and Lys336 (helix3) of RORγ. RESULTS |
|
59 72 charged clamp structure_element This interaction is further stabilized through a conserved charged clamp wherein the backbone amide of Tyr7 and carbonyl of Leu11 of EBI96 form hydrogen bonds with Glu504 (helix12) and Lys336 (helix3) of RORγ. RESULTS |
|
103 107 Tyr7 residue_name_number This interaction is further stabilized through a conserved charged clamp wherein the backbone amide of Tyr7 and carbonyl of Leu11 of EBI96 form hydrogen bonds with Glu504 (helix12) and Lys336 (helix3) of RORγ. RESULTS |
|
124 129 Leu11 residue_name_number This interaction is further stabilized through a conserved charged clamp wherein the backbone amide of Tyr7 and carbonyl of Leu11 of EBI96 form hydrogen bonds with Glu504 (helix12) and Lys336 (helix3) of RORγ. RESULTS |
|
133 138 EBI96 chemical This interaction is further stabilized through a conserved charged clamp wherein the backbone amide of Tyr7 and carbonyl of Leu11 of EBI96 form hydrogen bonds with Glu504 (helix12) and Lys336 (helix3) of RORγ. RESULTS |
|
144 158 hydrogen bonds bond_interaction This interaction is further stabilized through a conserved charged clamp wherein the backbone amide of Tyr7 and carbonyl of Leu11 of EBI96 form hydrogen bonds with Glu504 (helix12) and Lys336 (helix3) of RORγ. RESULTS |
|
164 170 Glu504 residue_name_number This interaction is further stabilized through a conserved charged clamp wherein the backbone amide of Tyr7 and carbonyl of Leu11 of EBI96 form hydrogen bonds with Glu504 (helix12) and Lys336 (helix3) of RORγ. RESULTS |
|
172 179 helix12 structure_element This interaction is further stabilized through a conserved charged clamp wherein the backbone amide of Tyr7 and carbonyl of Leu11 of EBI96 form hydrogen bonds with Glu504 (helix12) and Lys336 (helix3) of RORγ. RESULTS |
|
185 191 Lys336 residue_name_number This interaction is further stabilized through a conserved charged clamp wherein the backbone amide of Tyr7 and carbonyl of Leu11 of EBI96 form hydrogen bonds with Glu504 (helix12) and Lys336 (helix3) of RORγ. RESULTS |
|
193 199 helix3 structure_element This interaction is further stabilized through a conserved charged clamp wherein the backbone amide of Tyr7 and carbonyl of Leu11 of EBI96 form hydrogen bonds with Glu504 (helix12) and Lys336 (helix3) of RORγ. RESULTS |
|
204 208 RORγ protein This interaction is further stabilized through a conserved charged clamp wherein the backbone amide of Tyr7 and carbonyl of Leu11 of EBI96 form hydrogen bonds with Glu504 (helix12) and Lys336 (helix3) of RORγ. RESULTS |
|
18 31 charged clamp structure_element Formation of this charged clamp is essential for RORγ’s function for playing a role in transcriptional activation and this has been corroborated through mutagenic studies in this region. RESULTS |
|
49 53 RORγ protein Formation of this charged clamp is essential for RORγ’s function for playing a role in transcriptional activation and this has been corroborated through mutagenic studies in this region. RESULTS |
|
153 170 mutagenic studies experimental_method Formation of this charged clamp is essential for RORγ’s function for playing a role in transcriptional activation and this has been corroborated through mutagenic studies in this region. RESULTS |
|
0 6 BIO592 chemical BIO592 binds in a collapsed conformation stabilizing the agonist conformation of RORγ RESULTS |
|
18 27 collapsed protein_state BIO592 binds in a collapsed conformation stabilizing the agonist conformation of RORγ RESULTS |
|
57 64 agonist protein_state BIO592 binds in a collapsed conformation stabilizing the agonist conformation of RORγ RESULTS |
|
81 85 RORγ protein BIO592 binds in a collapsed conformation stabilizing the agonist conformation of RORγ RESULTS |
|
29 36 agonist protein_state a Collapsed binding mode of agonist BIO592 in the hydrophobic LBS of RORγ. FIG |
|
37 43 BIO592 chemical a Collapsed binding mode of agonist BIO592 in the hydrophobic LBS of RORγ. FIG |
|
63 66 LBS site a Collapsed binding mode of agonist BIO592 in the hydrophobic LBS of RORγ. FIG |
|
70 74 RORγ protein a Collapsed binding mode of agonist BIO592 in the hydrophobic LBS of RORγ. FIG |
|
2 15 Benzoxazinone chemical b Benzoxazinone ring system of agonist BIO592 packing against His479 of RORγ stabilizing agonist conformation of the AF2 helix FIG |
|
31 38 agonist protein_state b Benzoxazinone ring system of agonist BIO592 packing against His479 of RORγ stabilizing agonist conformation of the AF2 helix FIG |
|
39 45 BIO592 chemical b Benzoxazinone ring system of agonist BIO592 packing against His479 of RORγ stabilizing agonist conformation of the AF2 helix FIG |
|
62 68 His479 residue_name_number b Benzoxazinone ring system of agonist BIO592 packing against His479 of RORγ stabilizing agonist conformation of the AF2 helix FIG |
|
72 76 RORγ protein b Benzoxazinone ring system of agonist BIO592 packing against His479 of RORγ stabilizing agonist conformation of the AF2 helix FIG |
|
89 96 agonist protein_state b Benzoxazinone ring system of agonist BIO592 packing against His479 of RORγ stabilizing agonist conformation of the AF2 helix FIG |
|
117 126 AF2 helix structure_element b Benzoxazinone ring system of agonist BIO592 packing against His479 of RORγ stabilizing agonist conformation of the AF2 helix FIG |
|
0 6 BIO592 chemical BIO592 bound in a collapsed conformational state in the LBS of RORγ with the xylene ring positioned at the bottom of the pocket making hydrophobic interactions with Val376, Phe378, Phe388 and Phe401, with the ethyl-benzoxazinone ring making several hydrophobic interactions with Trp317, Leu324, Met358, Leu391, Ile 400 and His479 (Fig. 3a, Additional file 3). RESULTS |
|
7 15 bound in protein_state BIO592 bound in a collapsed conformational state in the LBS of RORγ with the xylene ring positioned at the bottom of the pocket making hydrophobic interactions with Val376, Phe378, Phe388 and Phe401, with the ethyl-benzoxazinone ring making several hydrophobic interactions with Trp317, Leu324, Met358, Leu391, Ile 400 and His479 (Fig. 3a, Additional file 3). RESULTS |
|
18 27 collapsed protein_state BIO592 bound in a collapsed conformational state in the LBS of RORγ with the xylene ring positioned at the bottom of the pocket making hydrophobic interactions with Val376, Phe378, Phe388 and Phe401, with the ethyl-benzoxazinone ring making several hydrophobic interactions with Trp317, Leu324, Met358, Leu391, Ile 400 and His479 (Fig. 3a, Additional file 3). RESULTS |
|
56 59 LBS site BIO592 bound in a collapsed conformational state in the LBS of RORγ with the xylene ring positioned at the bottom of the pocket making hydrophobic interactions with Val376, Phe378, Phe388 and Phe401, with the ethyl-benzoxazinone ring making several hydrophobic interactions with Trp317, Leu324, Met358, Leu391, Ile 400 and His479 (Fig. 3a, Additional file 3). RESULTS |
|
63 67 RORγ protein BIO592 bound in a collapsed conformational state in the LBS of RORγ with the xylene ring positioned at the bottom of the pocket making hydrophobic interactions with Val376, Phe378, Phe388 and Phe401, with the ethyl-benzoxazinone ring making several hydrophobic interactions with Trp317, Leu324, Met358, Leu391, Ile 400 and His479 (Fig. 3a, Additional file 3). RESULTS |
|
77 83 xylene chemical BIO592 bound in a collapsed conformational state in the LBS of RORγ with the xylene ring positioned at the bottom of the pocket making hydrophobic interactions with Val376, Phe378, Phe388 and Phe401, with the ethyl-benzoxazinone ring making several hydrophobic interactions with Trp317, Leu324, Met358, Leu391, Ile 400 and His479 (Fig. 3a, Additional file 3). RESULTS |
|
121 127 pocket site BIO592 bound in a collapsed conformational state in the LBS of RORγ with the xylene ring positioned at the bottom of the pocket making hydrophobic interactions with Val376, Phe378, Phe388 and Phe401, with the ethyl-benzoxazinone ring making several hydrophobic interactions with Trp317, Leu324, Met358, Leu391, Ile 400 and His479 (Fig. 3a, Additional file 3). RESULTS |
|
135 159 hydrophobic interactions bond_interaction BIO592 bound in a collapsed conformational state in the LBS of RORγ with the xylene ring positioned at the bottom of the pocket making hydrophobic interactions with Val376, Phe378, Phe388 and Phe401, with the ethyl-benzoxazinone ring making several hydrophobic interactions with Trp317, Leu324, Met358, Leu391, Ile 400 and His479 (Fig. 3a, Additional file 3). RESULTS |
|
165 171 Val376 residue_name_number BIO592 bound in a collapsed conformational state in the LBS of RORγ with the xylene ring positioned at the bottom of the pocket making hydrophobic interactions with Val376, Phe378, Phe388 and Phe401, with the ethyl-benzoxazinone ring making several hydrophobic interactions with Trp317, Leu324, Met358, Leu391, Ile 400 and His479 (Fig. 3a, Additional file 3). RESULTS |
|
173 179 Phe378 residue_name_number BIO592 bound in a collapsed conformational state in the LBS of RORγ with the xylene ring positioned at the bottom of the pocket making hydrophobic interactions with Val376, Phe378, Phe388 and Phe401, with the ethyl-benzoxazinone ring making several hydrophobic interactions with Trp317, Leu324, Met358, Leu391, Ile 400 and His479 (Fig. 3a, Additional file 3). RESULTS |
|
181 187 Phe388 residue_name_number BIO592 bound in a collapsed conformational state in the LBS of RORγ with the xylene ring positioned at the bottom of the pocket making hydrophobic interactions with Val376, Phe378, Phe388 and Phe401, with the ethyl-benzoxazinone ring making several hydrophobic interactions with Trp317, Leu324, Met358, Leu391, Ile 400 and His479 (Fig. 3a, Additional file 3). RESULTS |
|
192 198 Phe401 residue_name_number BIO592 bound in a collapsed conformational state in the LBS of RORγ with the xylene ring positioned at the bottom of the pocket making hydrophobic interactions with Val376, Phe378, Phe388 and Phe401, with the ethyl-benzoxazinone ring making several hydrophobic interactions with Trp317, Leu324, Met358, Leu391, Ile 400 and His479 (Fig. 3a, Additional file 3). RESULTS |
|
209 228 ethyl-benzoxazinone chemical BIO592 bound in a collapsed conformational state in the LBS of RORγ with the xylene ring positioned at the bottom of the pocket making hydrophobic interactions with Val376, Phe378, Phe388 and Phe401, with the ethyl-benzoxazinone ring making several hydrophobic interactions with Trp317, Leu324, Met358, Leu391, Ile 400 and His479 (Fig. 3a, Additional file 3). RESULTS |
|
249 273 hydrophobic interactions bond_interaction BIO592 bound in a collapsed conformational state in the LBS of RORγ with the xylene ring positioned at the bottom of the pocket making hydrophobic interactions with Val376, Phe378, Phe388 and Phe401, with the ethyl-benzoxazinone ring making several hydrophobic interactions with Trp317, Leu324, Met358, Leu391, Ile 400 and His479 (Fig. 3a, Additional file 3). RESULTS |
|
279 285 Trp317 residue_name_number BIO592 bound in a collapsed conformational state in the LBS of RORγ with the xylene ring positioned at the bottom of the pocket making hydrophobic interactions with Val376, Phe378, Phe388 and Phe401, with the ethyl-benzoxazinone ring making several hydrophobic interactions with Trp317, Leu324, Met358, Leu391, Ile 400 and His479 (Fig. 3a, Additional file 3). RESULTS |
|
287 293 Leu324 residue_name_number BIO592 bound in a collapsed conformational state in the LBS of RORγ with the xylene ring positioned at the bottom of the pocket making hydrophobic interactions with Val376, Phe378, Phe388 and Phe401, with the ethyl-benzoxazinone ring making several hydrophobic interactions with Trp317, Leu324, Met358, Leu391, Ile 400 and His479 (Fig. 3a, Additional file 3). RESULTS |
|
295 301 Met358 residue_name_number BIO592 bound in a collapsed conformational state in the LBS of RORγ with the xylene ring positioned at the bottom of the pocket making hydrophobic interactions with Val376, Phe378, Phe388 and Phe401, with the ethyl-benzoxazinone ring making several hydrophobic interactions with Trp317, Leu324, Met358, Leu391, Ile 400 and His479 (Fig. 3a, Additional file 3). RESULTS |
|
303 309 Leu391 residue_name_number BIO592 bound in a collapsed conformational state in the LBS of RORγ with the xylene ring positioned at the bottom of the pocket making hydrophobic interactions with Val376, Phe378, Phe388 and Phe401, with the ethyl-benzoxazinone ring making several hydrophobic interactions with Trp317, Leu324, Met358, Leu391, Ile 400 and His479 (Fig. 3a, Additional file 3). RESULTS |
|
311 318 Ile 400 residue_name_number BIO592 bound in a collapsed conformational state in the LBS of RORγ with the xylene ring positioned at the bottom of the pocket making hydrophobic interactions with Val376, Phe378, Phe388 and Phe401, with the ethyl-benzoxazinone ring making several hydrophobic interactions with Trp317, Leu324, Met358, Leu391, Ile 400 and His479 (Fig. 3a, Additional file 3). RESULTS |
|
323 329 His479 residue_name_number BIO592 bound in a collapsed conformational state in the LBS of RORγ with the xylene ring positioned at the bottom of the pocket making hydrophobic interactions with Val376, Phe378, Phe388 and Phe401, with the ethyl-benzoxazinone ring making several hydrophobic interactions with Trp317, Leu324, Met358, Leu391, Ile 400 and His479 (Fig. 3a, Additional file 3). RESULTS |
|
4 12 sulfonyl chemical The sulfonyl group faces the entrance of the pocket, while the CF3 makes a hydrophobic contact with Ala327. RESULTS |
|
45 51 pocket site The sulfonyl group faces the entrance of the pocket, while the CF3 makes a hydrophobic contact with Ala327. RESULTS |
|
75 94 hydrophobic contact bond_interaction The sulfonyl group faces the entrance of the pocket, while the CF3 makes a hydrophobic contact with Ala327. RESULTS |
|
100 106 Ala327 residue_name_number The sulfonyl group faces the entrance of the pocket, while the CF3 makes a hydrophobic contact with Ala327. RESULTS |
|
0 23 Hydrophobic interaction bond_interaction Hydrophobic interaction between the ethyl group of the benzoxazinone and His479 reinforce the His479 sidechain position for making the hydrogen bond with Tyr502 thereby stabilizing the agonist conformation (Fig. 3b). RESULTS |
|
55 68 benzoxazinone chemical Hydrophobic interaction between the ethyl group of the benzoxazinone and His479 reinforce the His479 sidechain position for making the hydrogen bond with Tyr502 thereby stabilizing the agonist conformation (Fig. 3b). RESULTS |
|
73 79 His479 residue_name_number Hydrophobic interaction between the ethyl group of the benzoxazinone and His479 reinforce the His479 sidechain position for making the hydrogen bond with Tyr502 thereby stabilizing the agonist conformation (Fig. 3b). RESULTS |
|
94 100 His479 residue_name_number Hydrophobic interaction between the ethyl group of the benzoxazinone and His479 reinforce the His479 sidechain position for making the hydrogen bond with Tyr502 thereby stabilizing the agonist conformation (Fig. 3b). RESULTS |
|
135 148 hydrogen bond bond_interaction Hydrophobic interaction between the ethyl group of the benzoxazinone and His479 reinforce the His479 sidechain position for making the hydrogen bond with Tyr502 thereby stabilizing the agonist conformation (Fig. 3b). RESULTS |
|
154 160 Tyr502 residue_name_number Hydrophobic interaction between the ethyl group of the benzoxazinone and His479 reinforce the His479 sidechain position for making the hydrogen bond with Tyr502 thereby stabilizing the agonist conformation (Fig. 3b). RESULTS |
|
185 192 agonist protein_state Hydrophobic interaction between the ethyl group of the benzoxazinone and His479 reinforce the His479 sidechain position for making the hydrogen bond with Tyr502 thereby stabilizing the agonist conformation (Fig. 3b). RESULTS |
|
0 4 RORγ protein RORγ AF2 helix is sensitive to proteolysis in the presence of Inverse Agonist BIO399 RESULTS |
|
5 14 AF2 helix structure_element RORγ AF2 helix is sensitive to proteolysis in the presence of Inverse Agonist BIO399 RESULTS |
|
50 61 presence of protein_state RORγ AF2 helix is sensitive to proteolysis in the presence of Inverse Agonist BIO399 RESULTS |
|
62 77 Inverse Agonist protein_state RORγ AF2 helix is sensitive to proteolysis in the presence of Inverse Agonist BIO399 RESULTS |
|
78 84 BIO399 chemical RORγ AF2 helix is sensitive to proteolysis in the presence of Inverse Agonist BIO399 RESULTS |
|
19 37 co-crystallization experimental_method Next, we attempted co-crystallization with the inverse agonist BIO399. RESULTS |
|
47 62 inverse agonist protein_state Next, we attempted co-crystallization with the inverse agonist BIO399. RESULTS |
|
63 69 BIO399 chemical Next, we attempted co-crystallization with the inverse agonist BIO399. RESULTS |
|
19 34 crystallization experimental_method However, extensive crystallization efforts with BIO399 and RORγ518 or other AF2 intact constructs did not produce crystals. RESULTS |
|
48 54 BIO399 chemical However, extensive crystallization efforts with BIO399 and RORγ518 or other AF2 intact constructs did not produce crystals. RESULTS |
|
59 66 RORγ518 protein However, extensive crystallization efforts with BIO399 and RORγ518 or other AF2 intact constructs did not produce crystals. RESULTS |
|
76 79 AF2 structure_element However, extensive crystallization efforts with BIO399 and RORγ518 or other AF2 intact constructs did not produce crystals. RESULTS |
|
80 86 intact protein_state However, extensive crystallization efforts with BIO399 and RORγ518 or other AF2 intact constructs did not produce crystals. RESULTS |
|
114 122 crystals evidence However, extensive crystallization efforts with BIO399 and RORγ518 or other AF2 intact constructs did not produce crystals. RESULTS |
|
25 32 RORγ518 protein We hypothesized that the RORγ518 coactivator peptide interaction in the FRET assay was disrupted upon BIO399 binding and that a conformational rearrangement of the AF2 helix could have occurred, hindering crystallization. RESULTS |
|
72 82 FRET assay experimental_method We hypothesized that the RORγ518 coactivator peptide interaction in the FRET assay was disrupted upon BIO399 binding and that a conformational rearrangement of the AF2 helix could have occurred, hindering crystallization. RESULTS |
|
102 108 BIO399 chemical We hypothesized that the RORγ518 coactivator peptide interaction in the FRET assay was disrupted upon BIO399 binding and that a conformational rearrangement of the AF2 helix could have occurred, hindering crystallization. RESULTS |
|
164 173 AF2 helix structure_element We hypothesized that the RORγ518 coactivator peptide interaction in the FRET assay was disrupted upon BIO399 binding and that a conformational rearrangement of the AF2 helix could have occurred, hindering crystallization. RESULTS |
|
205 220 crystallization experimental_method We hypothesized that the RORγ518 coactivator peptide interaction in the FRET assay was disrupted upon BIO399 binding and that a conformational rearrangement of the AF2 helix could have occurred, hindering crystallization. RESULTS |
|
34 41 RORγ518 protein Specific proteolytic positions on RORγ518 when treated with Actinase E alone (Green) or in the presence of BIO399 (Red) and shared proteolytic sites (Yellow) FIG |
|
47 59 treated with experimental_method Specific proteolytic positions on RORγ518 when treated with Actinase E alone (Green) or in the presence of BIO399 (Red) and shared proteolytic sites (Yellow) FIG |
|
60 70 Actinase E protein Specific proteolytic positions on RORγ518 when treated with Actinase E alone (Green) or in the presence of BIO399 (Red) and shared proteolytic sites (Yellow) FIG |
|
95 106 presence of protein_state Specific proteolytic positions on RORγ518 when treated with Actinase E alone (Green) or in the presence of BIO399 (Red) and shared proteolytic sites (Yellow) FIG |
|
107 113 BIO399 chemical Specific proteolytic positions on RORγ518 when treated with Actinase E alone (Green) or in the presence of BIO399 (Red) and shared proteolytic sites (Yellow) FIG |
|
131 148 proteolytic sites site Specific proteolytic positions on RORγ518 when treated with Actinase E alone (Green) or in the presence of BIO399 (Red) and shared proteolytic sites (Yellow) FIG |
|
21 30 AF2 helix structure_element The unfolding of the AF2 helix has been observed for other nuclear hormone receptors when bound to an inverse agonist or antagonist. RESULTS |
|
59 84 nuclear hormone receptors protein_type The unfolding of the AF2 helix has been observed for other nuclear hormone receptors when bound to an inverse agonist or antagonist. RESULTS |
|
90 98 bound to protein_state The unfolding of the AF2 helix has been observed for other nuclear hormone receptors when bound to an inverse agonist or antagonist. RESULTS |
|
102 117 inverse agonist protein_state The unfolding of the AF2 helix has been observed for other nuclear hormone receptors when bound to an inverse agonist or antagonist. RESULTS |
|
8 27 partial proteolysis experimental_method We used partial proteolysis in combination with mass spectrometry to determine if BIO399 was causing the AF2 helix to unfold. RESULTS |
|
48 65 mass spectrometry experimental_method We used partial proteolysis in combination with mass spectrometry to determine if BIO399 was causing the AF2 helix to unfold. RESULTS |
|
82 88 BIO399 chemical We used partial proteolysis in combination with mass spectrometry to determine if BIO399 was causing the AF2 helix to unfold. RESULTS |
|
105 114 AF2 helix structure_element We used partial proteolysis in combination with mass spectrometry to determine if BIO399 was causing the AF2 helix to unfold. RESULTS |
|
15 37 Actinase E proteolysis experimental_method Results of the Actinase E proteolysis experiments on RORγ518, the ternary complex of RORγ518 with agonist BIO592 and coactivator EBI96, or in the presence of inverse agonist BIO399 supported our hypothesis. RESULTS |
|
53 60 RORγ518 protein Results of the Actinase E proteolysis experiments on RORγ518, the ternary complex of RORγ518 with agonist BIO592 and coactivator EBI96, or in the presence of inverse agonist BIO399 supported our hypothesis. RESULTS |
|
85 92 RORγ518 protein Results of the Actinase E proteolysis experiments on RORγ518, the ternary complex of RORγ518 with agonist BIO592 and coactivator EBI96, or in the presence of inverse agonist BIO399 supported our hypothesis. RESULTS |
|
98 105 agonist protein_state Results of the Actinase E proteolysis experiments on RORγ518, the ternary complex of RORγ518 with agonist BIO592 and coactivator EBI96, or in the presence of inverse agonist BIO399 supported our hypothesis. RESULTS |
|
106 112 BIO592 chemical Results of the Actinase E proteolysis experiments on RORγ518, the ternary complex of RORγ518 with agonist BIO592 and coactivator EBI96, or in the presence of inverse agonist BIO399 supported our hypothesis. RESULTS |
|
129 134 EBI96 chemical Results of the Actinase E proteolysis experiments on RORγ518, the ternary complex of RORγ518 with agonist BIO592 and coactivator EBI96, or in the presence of inverse agonist BIO399 supported our hypothesis. RESULTS |
|
146 157 presence of protein_state Results of the Actinase E proteolysis experiments on RORγ518, the ternary complex of RORγ518 with agonist BIO592 and coactivator EBI96, or in the presence of inverse agonist BIO399 supported our hypothesis. RESULTS |
|
158 173 inverse agonist protein_state Results of the Actinase E proteolysis experiments on RORγ518, the ternary complex of RORγ518 with agonist BIO592 and coactivator EBI96, or in the presence of inverse agonist BIO399 supported our hypothesis. RESULTS |
|
174 180 BIO399 chemical Results of the Actinase E proteolysis experiments on RORγ518, the ternary complex of RORγ518 with agonist BIO592 and coactivator EBI96, or in the presence of inverse agonist BIO399 supported our hypothesis. RESULTS |
|
16 37 fragmentation pattern evidence Analysis of the fragmentation pattern showed minimal proteolytic removal of the AF2 helix by Actinase E on RORγ518 alone (ending at 504 to 506) and the ternary complex remained primarily intact (ending at 515/518) (Additional file 4). RESULTS |
|
80 89 AF2 helix structure_element Analysis of the fragmentation pattern showed minimal proteolytic removal of the AF2 helix by Actinase E on RORγ518 alone (ending at 504 to 506) and the ternary complex remained primarily intact (ending at 515/518) (Additional file 4). RESULTS |
|
93 103 Actinase E protein Analysis of the fragmentation pattern showed minimal proteolytic removal of the AF2 helix by Actinase E on RORγ518 alone (ending at 504 to 506) and the ternary complex remained primarily intact (ending at 515/518) (Additional file 4). RESULTS |
|
107 114 RORγ518 protein Analysis of the fragmentation pattern showed minimal proteolytic removal of the AF2 helix by Actinase E on RORγ518 alone (ending at 504 to 506) and the ternary complex remained primarily intact (ending at 515/518) (Additional file 4). RESULTS |
|
132 142 504 to 506 residue_range Analysis of the fragmentation pattern showed minimal proteolytic removal of the AF2 helix by Actinase E on RORγ518 alone (ending at 504 to 506) and the ternary complex remained primarily intact (ending at 515/518) (Additional file 4). RESULTS |
|
152 167 ternary complex protein_state Analysis of the fragmentation pattern showed minimal proteolytic removal of the AF2 helix by Actinase E on RORγ518 alone (ending at 504 to 506) and the ternary complex remained primarily intact (ending at 515/518) (Additional file 4). RESULTS |
|
205 208 515 residue_number Analysis of the fragmentation pattern showed minimal proteolytic removal of the AF2 helix by Actinase E on RORγ518 alone (ending at 504 to 506) and the ternary complex remained primarily intact (ending at 515/518) (Additional file 4). RESULTS |
|
209 212 518 residue_number Analysis of the fragmentation pattern showed minimal proteolytic removal of the AF2 helix by Actinase E on RORγ518 alone (ending at 504 to 506) and the ternary complex remained primarily intact (ending at 515/518) (Additional file 4). RESULTS |
|
16 27 presence of protein_state However, in the presence of inverse agonist BIO399, the proteolytic pattern showed significantly less protection, albeit the products were more heterogeneous (majority ending at 494/495), indicating the destabilization of the AF2 helix compared to either the APO or ternary agonist complex (Fig. 4, Additional file 5). RESULTS |
|
28 43 inverse agonist protein_state However, in the presence of inverse agonist BIO399, the proteolytic pattern showed significantly less protection, albeit the products were more heterogeneous (majority ending at 494/495), indicating the destabilization of the AF2 helix compared to either the APO or ternary agonist complex (Fig. 4, Additional file 5). RESULTS |
|
44 50 BIO399 chemical However, in the presence of inverse agonist BIO399, the proteolytic pattern showed significantly less protection, albeit the products were more heterogeneous (majority ending at 494/495), indicating the destabilization of the AF2 helix compared to either the APO or ternary agonist complex (Fig. 4, Additional file 5). RESULTS |
|
56 75 proteolytic pattern evidence However, in the presence of inverse agonist BIO399, the proteolytic pattern showed significantly less protection, albeit the products were more heterogeneous (majority ending at 494/495), indicating the destabilization of the AF2 helix compared to either the APO or ternary agonist complex (Fig. 4, Additional file 5). RESULTS |
|
178 181 494 residue_number However, in the presence of inverse agonist BIO399, the proteolytic pattern showed significantly less protection, albeit the products were more heterogeneous (majority ending at 494/495), indicating the destabilization of the AF2 helix compared to either the APO or ternary agonist complex (Fig. 4, Additional file 5). RESULTS |
|
182 185 495 residue_number However, in the presence of inverse agonist BIO399, the proteolytic pattern showed significantly less protection, albeit the products were more heterogeneous (majority ending at 494/495), indicating the destabilization of the AF2 helix compared to either the APO or ternary agonist complex (Fig. 4, Additional file 5). RESULTS |
|
226 235 AF2 helix structure_element However, in the presence of inverse agonist BIO399, the proteolytic pattern showed significantly less protection, albeit the products were more heterogeneous (majority ending at 494/495), indicating the destabilization of the AF2 helix compared to either the APO or ternary agonist complex (Fig. 4, Additional file 5). RESULTS |
|
259 262 APO protein_state However, in the presence of inverse agonist BIO399, the proteolytic pattern showed significantly less protection, albeit the products were more heterogeneous (majority ending at 494/495), indicating the destabilization of the AF2 helix compared to either the APO or ternary agonist complex (Fig. 4, Additional file 5). RESULTS |
|
266 289 ternary agonist complex protein_state However, in the presence of inverse agonist BIO399, the proteolytic pattern showed significantly less protection, albeit the products were more heterogeneous (majority ending at 494/495), indicating the destabilization of the AF2 helix compared to either the APO or ternary agonist complex (Fig. 4, Additional file 5). RESULTS |
|
18 35 cocrystallization experimental_method Several rounds of cocrystallization attempts with RORγ518 or other RORγ AF2 helix containing constructs complexed with BIO399 had not produced crystals. RESULTS |
|
50 57 RORγ518 protein Several rounds of cocrystallization attempts with RORγ518 or other RORγ AF2 helix containing constructs complexed with BIO399 had not produced crystals. RESULTS |
|
67 71 RORγ protein Several rounds of cocrystallization attempts with RORγ518 or other RORγ AF2 helix containing constructs complexed with BIO399 had not produced crystals. RESULTS |
|
72 81 AF2 helix structure_element Several rounds of cocrystallization attempts with RORγ518 or other RORγ AF2 helix containing constructs complexed with BIO399 had not produced crystals. RESULTS |
|
104 118 complexed with protein_state Several rounds of cocrystallization attempts with RORγ518 or other RORγ AF2 helix containing constructs complexed with BIO399 had not produced crystals. RESULTS |
|
119 125 BIO399 chemical Several rounds of cocrystallization attempts with RORγ518 or other RORγ AF2 helix containing constructs complexed with BIO399 had not produced crystals. RESULTS |
|
143 151 crystals evidence Several rounds of cocrystallization attempts with RORγ518 or other RORγ AF2 helix containing constructs complexed with BIO399 had not produced crystals. RESULTS |
|
36 44 crystals evidence We attributed the inability to form crystals to the unfolding of the AF2 helix induced by BIO399. RESULTS |
|
69 78 AF2 helix structure_element We attributed the inability to form crystals to the unfolding of the AF2 helix induced by BIO399. RESULTS |
|
90 96 BIO399 chemical We attributed the inability to form crystals to the unfolding of the AF2 helix induced by BIO399. RESULTS |
|
40 48 unfolded protein_state We reasoned that if we could remove the unfolded AF2 helix using proteolysis we could produce a binary complex more amenable to crystallization. RESULTS |
|
49 58 AF2 helix structure_element We reasoned that if we could remove the unfolded AF2 helix using proteolysis we could produce a binary complex more amenable to crystallization. RESULTS |
|
65 76 proteolysis experimental_method We reasoned that if we could remove the unfolded AF2 helix using proteolysis we could produce a binary complex more amenable to crystallization. RESULTS |
|
128 143 crystallization experimental_method We reasoned that if we could remove the unfolded AF2 helix using proteolysis we could produce a binary complex more amenable to crystallization. RESULTS |
|
0 13 AF2 truncated protein_state AF2 truncated RORγ BIO399 complex is more amenable to crystallization RESULTS |
|
14 25 RORγ BIO399 complex_assembly AF2 truncated RORγ BIO399 complex is more amenable to crystallization RESULTS |
|
54 69 crystallization experimental_method AF2 truncated RORγ BIO399 complex is more amenable to crystallization RESULTS |
|
14 23 structure evidence a The binary structure of AF2-truncated RORγ and BIO399. FIG |
|
27 40 AF2-truncated protein_state a The binary structure of AF2-truncated RORγ and BIO399. FIG |
|
41 45 RORγ protein a The binary structure of AF2-truncated RORγ and BIO399. FIG |
|
50 56 BIO399 chemical a The binary structure of AF2-truncated RORγ and BIO399. FIG |
|
6 19 superposition experimental_method b The superposition of inverse agonist BIO399 (Cyan) and agonist BIO592 (Green). FIG |
|
23 38 inverse agonist protein_state b The superposition of inverse agonist BIO399 (Cyan) and agonist BIO592 (Green). FIG |
|
39 45 BIO399 chemical b The superposition of inverse agonist BIO399 (Cyan) and agonist BIO592 (Green). FIG |
|
57 64 agonist protein_state b The superposition of inverse agonist BIO399 (Cyan) and agonist BIO592 (Green). FIG |
|
65 71 BIO592 chemical b The superposition of inverse agonist BIO399 (Cyan) and agonist BIO592 (Green). FIG |
|
14 20 Met358 residue_name_number c Movement of Met358 and His479 in the BIO399 (Cyan) and BIO592 (Green) structures FIG |
|
25 31 His479 residue_name_number c Movement of Met358 and His479 in the BIO399 (Cyan) and BIO592 (Green) structures FIG |
|
39 45 BIO399 chemical c Movement of Met358 and His479 in the BIO399 (Cyan) and BIO592 (Green) structures FIG |
|
57 63 BIO592 chemical c Movement of Met358 and His479 in the BIO399 (Cyan) and BIO592 (Green) structures FIG |
|
72 82 structures evidence c Movement of Met358 and His479 in the BIO399 (Cyan) and BIO592 (Green) structures FIG |
|
4 14 Actinase E protein The Actinase E treated RORγ518 BIO399 ternary complex (aeRORγ493/4) co-crystallized readily in several PEG based conditions. RESULTS |
|
23 37 RORγ518 BIO399 complex_assembly The Actinase E treated RORγ518 BIO399 ternary complex (aeRORγ493/4) co-crystallized readily in several PEG based conditions. RESULTS |
|
55 66 aeRORγ493/4 complex_assembly The Actinase E treated RORγ518 BIO399 ternary complex (aeRORγ493/4) co-crystallized readily in several PEG based conditions. RESULTS |
|
68 83 co-crystallized experimental_method The Actinase E treated RORγ518 BIO399 ternary complex (aeRORγ493/4) co-crystallized readily in several PEG based conditions. RESULTS |
|
4 13 structure evidence The structure of aeRORγ493/4 BIO399 complex was solved to 2.3 Å and adopted a similar core fold to the BIO592 agonist crystal structure (Fig. 5a, Additional file 3). RESULTS |
|
17 35 aeRORγ493/4 BIO399 complex_assembly The structure of aeRORγ493/4 BIO399 complex was solved to 2.3 Å and adopted a similar core fold to the BIO592 agonist crystal structure (Fig. 5a, Additional file 3). RESULTS |
|
48 54 solved experimental_method The structure of aeRORγ493/4 BIO399 complex was solved to 2.3 Å and adopted a similar core fold to the BIO592 agonist crystal structure (Fig. 5a, Additional file 3). RESULTS |
|
103 109 BIO592 chemical The structure of aeRORγ493/4 BIO399 complex was solved to 2.3 Å and adopted a similar core fold to the BIO592 agonist crystal structure (Fig. 5a, Additional file 3). RESULTS |
|
110 117 agonist protein_state The structure of aeRORγ493/4 BIO399 complex was solved to 2.3 Å and adopted a similar core fold to the BIO592 agonist crystal structure (Fig. 5a, Additional file 3). RESULTS |
|
118 135 crystal structure evidence The structure of aeRORγ493/4 BIO399 complex was solved to 2.3 Å and adopted a similar core fold to the BIO592 agonist crystal structure (Fig. 5a, Additional file 3). RESULTS |
|
4 22 aeRORγ493/4 BIO399 complex_assembly The aeRORγ493/4 BIO399 structure diverged at the c-terminal end of Helix 11 from the RORγ518 BIO592 EBI96 structure, where helix 11 unwinds into a random coil after residue L475. RESULTS |
|
23 32 structure evidence The aeRORγ493/4 BIO399 structure diverged at the c-terminal end of Helix 11 from the RORγ518 BIO592 EBI96 structure, where helix 11 unwinds into a random coil after residue L475. RESULTS |
|
67 75 Helix 11 structure_element The aeRORγ493/4 BIO399 structure diverged at the c-terminal end of Helix 11 from the RORγ518 BIO592 EBI96 structure, where helix 11 unwinds into a random coil after residue L475. RESULTS |
|
85 105 RORγ518 BIO592 EBI96 complex_assembly The aeRORγ493/4 BIO399 structure diverged at the c-terminal end of Helix 11 from the RORγ518 BIO592 EBI96 structure, where helix 11 unwinds into a random coil after residue L475. RESULTS |
|
106 115 structure evidence The aeRORγ493/4 BIO399 structure diverged at the c-terminal end of Helix 11 from the RORγ518 BIO592 EBI96 structure, where helix 11 unwinds into a random coil after residue L475. RESULTS |
|
123 131 helix 11 structure_element The aeRORγ493/4 BIO399 structure diverged at the c-terminal end of Helix 11 from the RORγ518 BIO592 EBI96 structure, where helix 11 unwinds into a random coil after residue L475. RESULTS |
|
173 177 L475 residue_name_number The aeRORγ493/4 BIO399 structure diverged at the c-terminal end of Helix 11 from the RORγ518 BIO592 EBI96 structure, where helix 11 unwinds into a random coil after residue L475. RESULTS |
|
0 15 Inverse agonist protein_state Inverse agonist BIO399 uses Met358 as a trigger for inverse agonism RESULTS |
|
16 22 BIO399 chemical Inverse agonist BIO399 uses Met358 as a trigger for inverse agonism RESULTS |
|
28 34 Met358 residue_name_number Inverse agonist BIO399 uses Met358 as a trigger for inverse agonism RESULTS |
|
0 6 BIO399 chemical BIO399 binds to the ligand binding site of RORγ adopting a collapsed conformation as seen with BIO592 where the two compounds superimpose with an RMSD of 0.72 Å (Fig. 5b). RESULTS |
|
20 39 ligand binding site site BIO399 binds to the ligand binding site of RORγ adopting a collapsed conformation as seen with BIO592 where the two compounds superimpose with an RMSD of 0.72 Å (Fig. 5b). RESULTS |
|
43 47 RORγ protein BIO399 binds to the ligand binding site of RORγ adopting a collapsed conformation as seen with BIO592 where the two compounds superimpose with an RMSD of 0.72 Å (Fig. 5b). RESULTS |
|
59 68 collapsed protein_state BIO399 binds to the ligand binding site of RORγ adopting a collapsed conformation as seen with BIO592 where the two compounds superimpose with an RMSD of 0.72 Å (Fig. 5b). RESULTS |
|
95 101 BIO592 chemical BIO399 binds to the ligand binding site of RORγ adopting a collapsed conformation as seen with BIO592 where the two compounds superimpose with an RMSD of 0.72 Å (Fig. 5b). RESULTS |
|
126 137 superimpose experimental_method BIO399 binds to the ligand binding site of RORγ adopting a collapsed conformation as seen with BIO592 where the two compounds superimpose with an RMSD of 0.72 Å (Fig. 5b). RESULTS |
|
146 150 RMSD evidence BIO399 binds to the ligand binding site of RORγ adopting a collapsed conformation as seen with BIO592 where the two compounds superimpose with an RMSD of 0.72 Å (Fig. 5b). RESULTS |
|
46 52 BIO399 chemical The majority of the side chains within 4 Å of BIO399 and BIO592 adopt similar rotomer conformations with the exceptions of Met358 and His479 (Fig. 5c). RESULTS |
|
57 63 BIO592 chemical The majority of the side chains within 4 Å of BIO399 and BIO592 adopt similar rotomer conformations with the exceptions of Met358 and His479 (Fig. 5c). RESULTS |
|
123 129 Met358 residue_name_number The majority of the side chains within 4 Å of BIO399 and BIO592 adopt similar rotomer conformations with the exceptions of Met358 and His479 (Fig. 5c). RESULTS |
|
134 140 His479 residue_name_number The majority of the side chains within 4 Å of BIO399 and BIO592 adopt similar rotomer conformations with the exceptions of Met358 and His479 (Fig. 5c). RESULTS |
|
4 26 difference density map evidence The difference density map showed clear positive density for Met358 in an alternate rotomer conformation compared to the one observed in the molecular replacement model or the other agonist containing models (Additional file 6). RESULTS |
|
40 56 positive density evidence The difference density map showed clear positive density for Met358 in an alternate rotomer conformation compared to the one observed in the molecular replacement model or the other agonist containing models (Additional file 6). RESULTS |
|
61 67 Met358 residue_name_number The difference density map showed clear positive density for Met358 in an alternate rotomer conformation compared to the one observed in the molecular replacement model or the other agonist containing models (Additional file 6). RESULTS |
|
141 168 molecular replacement model experimental_method The difference density map showed clear positive density for Met358 in an alternate rotomer conformation compared to the one observed in the molecular replacement model or the other agonist containing models (Additional file 6). RESULTS |
|
182 189 agonist protein_state The difference density map showed clear positive density for Met358 in an alternate rotomer conformation compared to the one observed in the molecular replacement model or the other agonist containing models (Additional file 6). RESULTS |
|
19 25 Met358 residue_name_number We tried to refine Met358 in the same conformation as the molecular replacement model or the other agonist containing models, but the results clearly indicated that this was not possible, thus confirming the new rotamer conformation for the Met358 sidechain in the inverse agonist bound structure. RESULTS |
|
58 85 molecular replacement model experimental_method We tried to refine Met358 in the same conformation as the molecular replacement model or the other agonist containing models, but the results clearly indicated that this was not possible, thus confirming the new rotamer conformation for the Met358 sidechain in the inverse agonist bound structure. RESULTS |
|
99 106 agonist protein_state We tried to refine Met358 in the same conformation as the molecular replacement model or the other agonist containing models, but the results clearly indicated that this was not possible, thus confirming the new rotamer conformation for the Met358 sidechain in the inverse agonist bound structure. RESULTS |
|
241 247 Met358 residue_name_number We tried to refine Met358 in the same conformation as the molecular replacement model or the other agonist containing models, but the results clearly indicated that this was not possible, thus confirming the new rotamer conformation for the Met358 sidechain in the inverse agonist bound structure. RESULTS |
|
265 286 inverse agonist bound protein_state We tried to refine Met358 in the same conformation as the molecular replacement model or the other agonist containing models, but the results clearly indicated that this was not possible, thus confirming the new rotamer conformation for the Met358 sidechain in the inverse agonist bound structure. RESULTS |
|
287 296 structure evidence We tried to refine Met358 in the same conformation as the molecular replacement model or the other agonist containing models, but the results clearly indicated that this was not possible, thus confirming the new rotamer conformation for the Met358 sidechain in the inverse agonist bound structure. RESULTS |
|
38 44 Met358 residue_name_number The change in rotomer conformation of Met358 between the agonist and inverse agonist structures is attributed to the gem-dimethyl group on the larger 7 membered benzoxazinone ring system of BIO399. RESULTS |
|
57 64 agonist protein_state The change in rotomer conformation of Met358 between the agonist and inverse agonist structures is attributed to the gem-dimethyl group on the larger 7 membered benzoxazinone ring system of BIO399. RESULTS |
|
69 84 inverse agonist protein_state The change in rotomer conformation of Met358 between the agonist and inverse agonist structures is attributed to the gem-dimethyl group on the larger 7 membered benzoxazinone ring system of BIO399. RESULTS |
|
85 95 structures evidence The change in rotomer conformation of Met358 between the agonist and inverse agonist structures is attributed to the gem-dimethyl group on the larger 7 membered benzoxazinone ring system of BIO399. RESULTS |
|
161 174 benzoxazinone chemical The change in rotomer conformation of Met358 between the agonist and inverse agonist structures is attributed to the gem-dimethyl group on the larger 7 membered benzoxazinone ring system of BIO399. RESULTS |
|
190 196 BIO399 chemical The change in rotomer conformation of Met358 between the agonist and inverse agonist structures is attributed to the gem-dimethyl group on the larger 7 membered benzoxazinone ring system of BIO399. RESULTS |
|
4 14 comparison experimental_method The comparison of the two structures shows that the agonist conformation observed in the BIO592 structure would be perturbed by BIO399 pushing Met358 into Phe506 of the AF2 helix indicating that Met358 is a trigger for inducing inverse agonism in RORγ (Fig. 5c). RESULTS |
|
26 36 structures evidence The comparison of the two structures shows that the agonist conformation observed in the BIO592 structure would be perturbed by BIO399 pushing Met358 into Phe506 of the AF2 helix indicating that Met358 is a trigger for inducing inverse agonism in RORγ (Fig. 5c). RESULTS |
|
52 59 agonist protein_state The comparison of the two structures shows that the agonist conformation observed in the BIO592 structure would be perturbed by BIO399 pushing Met358 into Phe506 of the AF2 helix indicating that Met358 is a trigger for inducing inverse agonism in RORγ (Fig. 5c). RESULTS |
|
89 95 BIO592 chemical The comparison of the two structures shows that the agonist conformation observed in the BIO592 structure would be perturbed by BIO399 pushing Met358 into Phe506 of the AF2 helix indicating that Met358 is a trigger for inducing inverse agonism in RORγ (Fig. 5c). RESULTS |
|
96 105 structure evidence The comparison of the two structures shows that the agonist conformation observed in the BIO592 structure would be perturbed by BIO399 pushing Met358 into Phe506 of the AF2 helix indicating that Met358 is a trigger for inducing inverse agonism in RORγ (Fig. 5c). RESULTS |
|
128 134 BIO399 chemical The comparison of the two structures shows that the agonist conformation observed in the BIO592 structure would be perturbed by BIO399 pushing Met358 into Phe506 of the AF2 helix indicating that Met358 is a trigger for inducing inverse agonism in RORγ (Fig. 5c). RESULTS |
|
143 149 Met358 residue_name_number The comparison of the two structures shows that the agonist conformation observed in the BIO592 structure would be perturbed by BIO399 pushing Met358 into Phe506 of the AF2 helix indicating that Met358 is a trigger for inducing inverse agonism in RORγ (Fig. 5c). RESULTS |
|
155 161 Phe506 residue_name_number The comparison of the two structures shows that the agonist conformation observed in the BIO592 structure would be perturbed by BIO399 pushing Met358 into Phe506 of the AF2 helix indicating that Met358 is a trigger for inducing inverse agonism in RORγ (Fig. 5c). RESULTS |
|
169 178 AF2 helix structure_element The comparison of the two structures shows that the agonist conformation observed in the BIO592 structure would be perturbed by BIO399 pushing Met358 into Phe506 of the AF2 helix indicating that Met358 is a trigger for inducing inverse agonism in RORγ (Fig. 5c). RESULTS |
|
195 201 Met358 residue_name_number The comparison of the two structures shows that the agonist conformation observed in the BIO592 structure would be perturbed by BIO399 pushing Met358 into Phe506 of the AF2 helix indicating that Met358 is a trigger for inducing inverse agonism in RORγ (Fig. 5c). RESULTS |
|
247 251 RORγ protein The comparison of the two structures shows that the agonist conformation observed in the BIO592 structure would be perturbed by BIO399 pushing Met358 into Phe506 of the AF2 helix indicating that Met358 is a trigger for inducing inverse agonism in RORγ (Fig. 5c). RESULTS |
|
0 6 BIO399 chemical BIO399 and Inverse agonist T0901317 bind in a collapsed conformation distinct from other RORγ Inverse Agonists Cocrystal structures RESULTS |
|
11 26 Inverse agonist protein_state BIO399 and Inverse agonist T0901317 bind in a collapsed conformation distinct from other RORγ Inverse Agonists Cocrystal structures RESULTS |
|
27 35 T0901317 chemical BIO399 and Inverse agonist T0901317 bind in a collapsed conformation distinct from other RORγ Inverse Agonists Cocrystal structures RESULTS |
|
46 55 collapsed protein_state BIO399 and Inverse agonist T0901317 bind in a collapsed conformation distinct from other RORγ Inverse Agonists Cocrystal structures RESULTS |
|
89 93 RORγ protein BIO399 and Inverse agonist T0901317 bind in a collapsed conformation distinct from other RORγ Inverse Agonists Cocrystal structures RESULTS |
|
111 131 Cocrystal structures evidence BIO399 and Inverse agonist T0901317 bind in a collapsed conformation distinct from other RORγ Inverse Agonists Cocrystal structures RESULTS |
|
3 10 Overlay experimental_method a Overlay of RORγ structures bound to BIO596 (Green), BIO399 (Cyan) and T0901317 (Pink). FIG |
|
14 18 RORγ protein a Overlay of RORγ structures bound to BIO596 (Green), BIO399 (Cyan) and T0901317 (Pink). FIG |
|
19 29 structures evidence a Overlay of RORγ structures bound to BIO596 (Green), BIO399 (Cyan) and T0901317 (Pink). FIG |
|
30 38 bound to protein_state a Overlay of RORγ structures bound to BIO596 (Green), BIO399 (Cyan) and T0901317 (Pink). FIG |
|
39 45 BIO596 chemical a Overlay of RORγ structures bound to BIO596 (Green), BIO399 (Cyan) and T0901317 (Pink). FIG |
|
55 61 BIO399 chemical a Overlay of RORγ structures bound to BIO596 (Green), BIO399 (Cyan) and T0901317 (Pink). FIG |
|
73 81 T0901317 chemical a Overlay of RORγ structures bound to BIO596 (Green), BIO399 (Cyan) and T0901317 (Pink). FIG |
|
2 9 Overlay experimental_method b Overlay of M358 in RORγ structure BIO596 (Green), BIO399 (Cyan), Digoxin (Yellow), Compound 2 (Grey), Compound 48 (Salmon) and Compound 4j (Orange) FIG |
|
13 17 M358 residue_name_number b Overlay of M358 in RORγ structure BIO596 (Green), BIO399 (Cyan), Digoxin (Yellow), Compound 2 (Grey), Compound 48 (Salmon) and Compound 4j (Orange) FIG |
|
21 25 RORγ protein b Overlay of M358 in RORγ structure BIO596 (Green), BIO399 (Cyan), Digoxin (Yellow), Compound 2 (Grey), Compound 48 (Salmon) and Compound 4j (Orange) FIG |
|
26 35 structure evidence b Overlay of M358 in RORγ structure BIO596 (Green), BIO399 (Cyan), Digoxin (Yellow), Compound 2 (Grey), Compound 48 (Salmon) and Compound 4j (Orange) FIG |
|
36 42 BIO596 chemical b Overlay of M358 in RORγ structure BIO596 (Green), BIO399 (Cyan), Digoxin (Yellow), Compound 2 (Grey), Compound 48 (Salmon) and Compound 4j (Orange) FIG |
|
52 58 BIO399 chemical b Overlay of M358 in RORγ structure BIO596 (Green), BIO399 (Cyan), Digoxin (Yellow), Compound 2 (Grey), Compound 48 (Salmon) and Compound 4j (Orange) FIG |
|
67 74 Digoxin chemical b Overlay of M358 in RORγ structure BIO596 (Green), BIO399 (Cyan), Digoxin (Yellow), Compound 2 (Grey), Compound 48 (Salmon) and Compound 4j (Orange) FIG |
|
4 24 co-crystal structure evidence The co-crystal structure of RORγ with T0901317 (PDB code: 4NB6), an inverse agonist of RORγ (IC50 of 54nM in an SRC1 displacement FRET assay and an IC50 of 59nM in our FRET assay (Additional file 7)) shows that it adopts a collapsed conformation similar to the structure of BIO399 described here. RESULTS |
|
28 32 RORγ protein The co-crystal structure of RORγ with T0901317 (PDB code: 4NB6), an inverse agonist of RORγ (IC50 of 54nM in an SRC1 displacement FRET assay and an IC50 of 59nM in our FRET assay (Additional file 7)) shows that it adopts a collapsed conformation similar to the structure of BIO399 described here. RESULTS |
|
38 46 T0901317 chemical The co-crystal structure of RORγ with T0901317 (PDB code: 4NB6), an inverse agonist of RORγ (IC50 of 54nM in an SRC1 displacement FRET assay and an IC50 of 59nM in our FRET assay (Additional file 7)) shows that it adopts a collapsed conformation similar to the structure of BIO399 described here. RESULTS |
|
68 83 inverse agonist protein_state The co-crystal structure of RORγ with T0901317 (PDB code: 4NB6), an inverse agonist of RORγ (IC50 of 54nM in an SRC1 displacement FRET assay and an IC50 of 59nM in our FRET assay (Additional file 7)) shows that it adopts a collapsed conformation similar to the structure of BIO399 described here. RESULTS |
|
87 91 RORγ protein The co-crystal structure of RORγ with T0901317 (PDB code: 4NB6), an inverse agonist of RORγ (IC50 of 54nM in an SRC1 displacement FRET assay and an IC50 of 59nM in our FRET assay (Additional file 7)) shows that it adopts a collapsed conformation similar to the structure of BIO399 described here. RESULTS |
|
93 97 IC50 evidence The co-crystal structure of RORγ with T0901317 (PDB code: 4NB6), an inverse agonist of RORγ (IC50 of 54nM in an SRC1 displacement FRET assay and an IC50 of 59nM in our FRET assay (Additional file 7)) shows that it adopts a collapsed conformation similar to the structure of BIO399 described here. RESULTS |
|
112 140 SRC1 displacement FRET assay experimental_method The co-crystal structure of RORγ with T0901317 (PDB code: 4NB6), an inverse agonist of RORγ (IC50 of 54nM in an SRC1 displacement FRET assay and an IC50 of 59nM in our FRET assay (Additional file 7)) shows that it adopts a collapsed conformation similar to the structure of BIO399 described here. RESULTS |
|
148 152 IC50 evidence The co-crystal structure of RORγ with T0901317 (PDB code: 4NB6), an inverse agonist of RORγ (IC50 of 54nM in an SRC1 displacement FRET assay and an IC50 of 59nM in our FRET assay (Additional file 7)) shows that it adopts a collapsed conformation similar to the structure of BIO399 described here. RESULTS |
|
168 178 FRET assay experimental_method The co-crystal structure of RORγ with T0901317 (PDB code: 4NB6), an inverse agonist of RORγ (IC50 of 54nM in an SRC1 displacement FRET assay and an IC50 of 59nM in our FRET assay (Additional file 7)) shows that it adopts a collapsed conformation similar to the structure of BIO399 described here. RESULTS |
|
223 232 collapsed protein_state The co-crystal structure of RORγ with T0901317 (PDB code: 4NB6), an inverse agonist of RORγ (IC50 of 54nM in an SRC1 displacement FRET assay and an IC50 of 59nM in our FRET assay (Additional file 7)) shows that it adopts a collapsed conformation similar to the structure of BIO399 described here. RESULTS |
|
261 270 structure evidence The co-crystal structure of RORγ with T0901317 (PDB code: 4NB6), an inverse agonist of RORγ (IC50 of 54nM in an SRC1 displacement FRET assay and an IC50 of 59nM in our FRET assay (Additional file 7)) shows that it adopts a collapsed conformation similar to the structure of BIO399 described here. RESULTS |
|
274 280 BIO399 chemical The co-crystal structure of RORγ with T0901317 (PDB code: 4NB6), an inverse agonist of RORγ (IC50 of 54nM in an SRC1 displacement FRET assay and an IC50 of 59nM in our FRET assay (Additional file 7)) shows that it adopts a collapsed conformation similar to the structure of BIO399 described here. RESULTS |
|
18 29 superimpose experimental_method The two compounds superimpose with an RMSD of 0.81 Å (Fig. 6a). RESULTS |
|
38 42 RMSD evidence The two compounds superimpose with an RMSD of 0.81 Å (Fig. 6a). RESULTS |
|
21 39 hexafluoropropanol chemical The CF3 group on the hexafluoropropanol group of T0901317 was reported to fit the electron density in two conformations one of which pushes Met358 into the vicinity of Phe506 in the RORγ BIO592 agonist structure. RESULTS |
|
49 57 T0901317 chemical The CF3 group on the hexafluoropropanol group of T0901317 was reported to fit the electron density in two conformations one of which pushes Met358 into the vicinity of Phe506 in the RORγ BIO592 agonist structure. RESULTS |
|
82 98 electron density evidence The CF3 group on the hexafluoropropanol group of T0901317 was reported to fit the electron density in two conformations one of which pushes Met358 into the vicinity of Phe506 in the RORγ BIO592 agonist structure. RESULTS |
|
140 146 Met358 residue_name_number The CF3 group on the hexafluoropropanol group of T0901317 was reported to fit the electron density in two conformations one of which pushes Met358 into the vicinity of Phe506 in the RORγ BIO592 agonist structure. RESULTS |
|
168 174 Phe506 residue_name_number The CF3 group on the hexafluoropropanol group of T0901317 was reported to fit the electron density in two conformations one of which pushes Met358 into the vicinity of Phe506 in the RORγ BIO592 agonist structure. RESULTS |
|
182 186 RORγ protein The CF3 group on the hexafluoropropanol group of T0901317 was reported to fit the electron density in two conformations one of which pushes Met358 into the vicinity of Phe506 in the RORγ BIO592 agonist structure. RESULTS |
|
187 193 BIO592 chemical The CF3 group on the hexafluoropropanol group of T0901317 was reported to fit the electron density in two conformations one of which pushes Met358 into the vicinity of Phe506 in the RORγ BIO592 agonist structure. RESULTS |
|
194 201 agonist protein_state The CF3 group on the hexafluoropropanol group of T0901317 was reported to fit the electron density in two conformations one of which pushes Met358 into the vicinity of Phe506 in the RORγ BIO592 agonist structure. RESULTS |
|
202 211 structure evidence The CF3 group on the hexafluoropropanol group of T0901317 was reported to fit the electron density in two conformations one of which pushes Met358 into the vicinity of Phe506 in the RORγ BIO592 agonist structure. RESULTS |
|
30 36 Met358 residue_name_number We hypothesize that since the Met358 sidechain conformation in the T0901317 RORγ structure is not in the BIO399 conformation, this difference could account for the 10-fold reduction in the inverse agonism for T0901317 compared to BIO399 in the FRET assay. RESULTS |
|
67 75 T0901317 chemical We hypothesize that since the Met358 sidechain conformation in the T0901317 RORγ structure is not in the BIO399 conformation, this difference could account for the 10-fold reduction in the inverse agonism for T0901317 compared to BIO399 in the FRET assay. RESULTS |
|
76 80 RORγ protein We hypothesize that since the Met358 sidechain conformation in the T0901317 RORγ structure is not in the BIO399 conformation, this difference could account for the 10-fold reduction in the inverse agonism for T0901317 compared to BIO399 in the FRET assay. RESULTS |
|
81 90 structure evidence We hypothesize that since the Met358 sidechain conformation in the T0901317 RORγ structure is not in the BIO399 conformation, this difference could account for the 10-fold reduction in the inverse agonism for T0901317 compared to BIO399 in the FRET assay. RESULTS |
|
105 111 BIO399 chemical We hypothesize that since the Met358 sidechain conformation in the T0901317 RORγ structure is not in the BIO399 conformation, this difference could account for the 10-fold reduction in the inverse agonism for T0901317 compared to BIO399 in the FRET assay. RESULTS |
|
209 217 T0901317 chemical We hypothesize that since the Met358 sidechain conformation in the T0901317 RORγ structure is not in the BIO399 conformation, this difference could account for the 10-fold reduction in the inverse agonism for T0901317 compared to BIO399 in the FRET assay. RESULTS |
|
230 236 BIO399 chemical We hypothesize that since the Met358 sidechain conformation in the T0901317 RORγ structure is not in the BIO399 conformation, this difference could account for the 10-fold reduction in the inverse agonism for T0901317 compared to BIO399 in the FRET assay. RESULTS |
|
244 254 FRET assay experimental_method We hypothesize that since the Met358 sidechain conformation in the T0901317 RORγ structure is not in the BIO399 conformation, this difference could account for the 10-fold reduction in the inverse agonism for T0901317 compared to BIO399 in the FRET assay. RESULTS |
|
0 21 Co-crystal structures evidence Co-crystal structures of RORγ have been generated with several potent inverse agonists adopting a linear conformation distinct from the collapsed conformations seen for BIO399 and T090131718. RESULTS |
|
25 29 RORγ protein Co-crystal structures of RORγ have been generated with several potent inverse agonists adopting a linear conformation distinct from the collapsed conformations seen for BIO399 and T090131718. RESULTS |
|
98 104 linear protein_state Co-crystal structures of RORγ have been generated with several potent inverse agonists adopting a linear conformation distinct from the collapsed conformations seen for BIO399 and T090131718. RESULTS |
|
136 145 collapsed protein_state Co-crystal structures of RORγ have been generated with several potent inverse agonists adopting a linear conformation distinct from the collapsed conformations seen for BIO399 and T090131718. RESULTS |
|
169 175 BIO399 chemical Co-crystal structures of RORγ have been generated with several potent inverse agonists adopting a linear conformation distinct from the collapsed conformations seen for BIO399 and T090131718. RESULTS |
|
180 190 T090131718 chemical Co-crystal structures of RORγ have been generated with several potent inverse agonists adopting a linear conformation distinct from the collapsed conformations seen for BIO399 and T090131718. RESULTS |
|
4 19 inverse agonist protein_state The inverse agonist activity for these compounds has been attributed to orientating Trp317 to clash with Tyr502 or a direct inverse agonist hydrogen bonding event with His479, both of which would perturb the agonist conformation of RORγ. RESULTS |
|
84 90 Trp317 residue_name_number The inverse agonist activity for these compounds has been attributed to orientating Trp317 to clash with Tyr502 or a direct inverse agonist hydrogen bonding event with His479, both of which would perturb the agonist conformation of RORγ. RESULTS |
|
105 111 Tyr502 residue_name_number The inverse agonist activity for these compounds has been attributed to orientating Trp317 to clash with Tyr502 or a direct inverse agonist hydrogen bonding event with His479, both of which would perturb the agonist conformation of RORγ. RESULTS |
|
124 139 inverse agonist protein_state The inverse agonist activity for these compounds has been attributed to orientating Trp317 to clash with Tyr502 or a direct inverse agonist hydrogen bonding event with His479, both of which would perturb the agonist conformation of RORγ. RESULTS |
|
140 156 hydrogen bonding bond_interaction The inverse agonist activity for these compounds has been attributed to orientating Trp317 to clash with Tyr502 or a direct inverse agonist hydrogen bonding event with His479, both of which would perturb the agonist conformation of RORγ. RESULTS |
|
168 174 His479 residue_name_number The inverse agonist activity for these compounds has been attributed to orientating Trp317 to clash with Tyr502 or a direct inverse agonist hydrogen bonding event with His479, both of which would perturb the agonist conformation of RORγ. RESULTS |
|
208 215 agonist protein_state The inverse agonist activity for these compounds has been attributed to orientating Trp317 to clash with Tyr502 or a direct inverse agonist hydrogen bonding event with His479, both of which would perturb the agonist conformation of RORγ. RESULTS |
|
232 236 RORγ protein The inverse agonist activity for these compounds has been attributed to orientating Trp317 to clash with Tyr502 or a direct inverse agonist hydrogen bonding event with His479, both of which would perturb the agonist conformation of RORγ. RESULTS |
|
0 6 BIO399 chemical BIO399 neither orients the sidechain of Trp317 toward Tyr502 nor forms a hydrogen bond with His479 suggesting its mode of action is distinct from linear inverse agonists (Additional file 8). RESULTS |
|
40 46 Trp317 residue_name_number BIO399 neither orients the sidechain of Trp317 toward Tyr502 nor forms a hydrogen bond with His479 suggesting its mode of action is distinct from linear inverse agonists (Additional file 8). RESULTS |
|
54 60 Tyr502 residue_name_number BIO399 neither orients the sidechain of Trp317 toward Tyr502 nor forms a hydrogen bond with His479 suggesting its mode of action is distinct from linear inverse agonists (Additional file 8). RESULTS |
|
73 86 hydrogen bond bond_interaction BIO399 neither orients the sidechain of Trp317 toward Tyr502 nor forms a hydrogen bond with His479 suggesting its mode of action is distinct from linear inverse agonists (Additional file 8). RESULTS |
|
92 98 His479 residue_name_number BIO399 neither orients the sidechain of Trp317 toward Tyr502 nor forms a hydrogen bond with His479 suggesting its mode of action is distinct from linear inverse agonists (Additional file 8). RESULTS |
|
14 29 inverse agonist protein_state In the linear inverse agonist crystal structures the side chain of Met358 resides in a similar position as the rotomer observed in RORγ agonist structures with BIO592 described here or as observed in the hydroxycholesterol derivatives and therefore would not trigger inverse agonism with these ligands (Fig. 6b). RESULTS |
|
30 48 crystal structures evidence In the linear inverse agonist crystal structures the side chain of Met358 resides in a similar position as the rotomer observed in RORγ agonist structures with BIO592 described here or as observed in the hydroxycholesterol derivatives and therefore would not trigger inverse agonism with these ligands (Fig. 6b). RESULTS |
|
67 73 Met358 residue_name_number In the linear inverse agonist crystal structures the side chain of Met358 resides in a similar position as the rotomer observed in RORγ agonist structures with BIO592 described here or as observed in the hydroxycholesterol derivatives and therefore would not trigger inverse agonism with these ligands (Fig. 6b). RESULTS |
|
131 135 RORγ protein In the linear inverse agonist crystal structures the side chain of Met358 resides in a similar position as the rotomer observed in RORγ agonist structures with BIO592 described here or as observed in the hydroxycholesterol derivatives and therefore would not trigger inverse agonism with these ligands (Fig. 6b). RESULTS |
|
136 143 agonist protein_state In the linear inverse agonist crystal structures the side chain of Met358 resides in a similar position as the rotomer observed in RORγ agonist structures with BIO592 described here or as observed in the hydroxycholesterol derivatives and therefore would not trigger inverse agonism with these ligands (Fig. 6b). RESULTS |
|
144 154 structures evidence In the linear inverse agonist crystal structures the side chain of Met358 resides in a similar position as the rotomer observed in RORγ agonist structures with BIO592 described here or as observed in the hydroxycholesterol derivatives and therefore would not trigger inverse agonism with these ligands (Fig. 6b). RESULTS |
|
160 166 BIO592 chemical In the linear inverse agonist crystal structures the side chain of Met358 resides in a similar position as the rotomer observed in RORγ agonist structures with BIO592 described here or as observed in the hydroxycholesterol derivatives and therefore would not trigger inverse agonism with these ligands (Fig. 6b). RESULTS |
|
204 222 hydroxycholesterol chemical In the linear inverse agonist crystal structures the side chain of Met358 resides in a similar position as the rotomer observed in RORγ agonist structures with BIO592 described here or as observed in the hydroxycholesterol derivatives and therefore would not trigger inverse agonism with these ligands (Fig. 6b). RESULTS |
|
0 6 BIO399 chemical BIO399 shows selectivity for RORγ over RORα and RORβ in a GAL4 Cellular Reporter Assay RESULTS |
|
29 33 RORγ protein BIO399 shows selectivity for RORγ over RORα and RORβ in a GAL4 Cellular Reporter Assay RESULTS |
|
39 43 RORα protein BIO399 shows selectivity for RORγ over RORα and RORβ in a GAL4 Cellular Reporter Assay RESULTS |
|
48 52 RORβ protein BIO399 shows selectivity for RORγ over RORα and RORβ in a GAL4 Cellular Reporter Assay RESULTS |
|
58 86 GAL4 Cellular Reporter Assay experimental_method BIO399 shows selectivity for RORγ over RORα and RORβ in a GAL4 Cellular Reporter Assay RESULTS |
|
0 15 GAL4 cell assay experimental_method GAL4 cell assay selectivity profile for BIO399 toward RORα and RORβ in GAL4 TABLE |
|
40 46 BIO399 chemical GAL4 cell assay selectivity profile for BIO399 toward RORα and RORβ in GAL4 TABLE |
|
54 58 RORα protein GAL4 cell assay selectivity profile for BIO399 toward RORα and RORβ in GAL4 TABLE |
|
63 67 RORβ protein GAL4 cell assay selectivity profile for BIO399 toward RORα and RORβ in GAL4 TABLE |
|
71 75 GAL4 protein GAL4 cell assay selectivity profile for BIO399 toward RORα and RORβ in GAL4 TABLE |
|
3 10 Overlay experimental_method a Overlay of RORα (yellow), β (pink) and γ (cyan) showing side chain differences at Met358 inverse agonism trigger position and (b) around the benzoxazinone ring system of BIO399 FIG |
|
14 18 RORα protein a Overlay of RORα (yellow), β (pink) and γ (cyan) showing side chain differences at Met358 inverse agonism trigger position and (b) around the benzoxazinone ring system of BIO399 FIG |
|
29 30 β protein a Overlay of RORα (yellow), β (pink) and γ (cyan) showing side chain differences at Met358 inverse agonism trigger position and (b) around the benzoxazinone ring system of BIO399 FIG |
|
42 43 γ protein a Overlay of RORα (yellow), β (pink) and γ (cyan) showing side chain differences at Met358 inverse agonism trigger position and (b) around the benzoxazinone ring system of BIO399 FIG |
|
85 91 Met358 residue_name_number a Overlay of RORα (yellow), β (pink) and γ (cyan) showing side chain differences at Met358 inverse agonism trigger position and (b) around the benzoxazinone ring system of BIO399 FIG |
|
144 157 benzoxazinone chemical a Overlay of RORα (yellow), β (pink) and γ (cyan) showing side chain differences at Met358 inverse agonism trigger position and (b) around the benzoxazinone ring system of BIO399 FIG |
|
173 179 BIO399 chemical a Overlay of RORα (yellow), β (pink) and γ (cyan) showing side chain differences at Met358 inverse agonism trigger position and (b) around the benzoxazinone ring system of BIO399 FIG |
|
54 60 BIO399 chemical In order to assess the in vivo selectivity profile of BIO399 a cellular reporter assay was implemented where the ligand binding domains of ROR α, β and γ were fused to the DNA binding domain of the transcriptional factor GAL4. RESULTS |
|
63 86 cellular reporter assay experimental_method In order to assess the in vivo selectivity profile of BIO399 a cellular reporter assay was implemented where the ligand binding domains of ROR α, β and γ were fused to the DNA binding domain of the transcriptional factor GAL4. RESULTS |
|
113 135 ligand binding domains structure_element In order to assess the in vivo selectivity profile of BIO399 a cellular reporter assay was implemented where the ligand binding domains of ROR α, β and γ were fused to the DNA binding domain of the transcriptional factor GAL4. RESULTS |
|
139 142 ROR protein_type In order to assess the in vivo selectivity profile of BIO399 a cellular reporter assay was implemented where the ligand binding domains of ROR α, β and γ were fused to the DNA binding domain of the transcriptional factor GAL4. RESULTS |
|
143 144 α protein In order to assess the in vivo selectivity profile of BIO399 a cellular reporter assay was implemented where the ligand binding domains of ROR α, β and γ were fused to the DNA binding domain of the transcriptional factor GAL4. RESULTS |
|
146 147 β protein In order to assess the in vivo selectivity profile of BIO399 a cellular reporter assay was implemented where the ligand binding domains of ROR α, β and γ were fused to the DNA binding domain of the transcriptional factor GAL4. RESULTS |
|
152 153 γ protein In order to assess the in vivo selectivity profile of BIO399 a cellular reporter assay was implemented where the ligand binding domains of ROR α, β and γ were fused to the DNA binding domain of the transcriptional factor GAL4. RESULTS |
|
159 167 fused to experimental_method In order to assess the in vivo selectivity profile of BIO399 a cellular reporter assay was implemented where the ligand binding domains of ROR α, β and γ were fused to the DNA binding domain of the transcriptional factor GAL4. RESULTS |
|
172 190 DNA binding domain structure_element In order to assess the in vivo selectivity profile of BIO399 a cellular reporter assay was implemented where the ligand binding domains of ROR α, β and γ were fused to the DNA binding domain of the transcriptional factor GAL4. RESULTS |
|
198 220 transcriptional factor protein_type In order to assess the in vivo selectivity profile of BIO399 a cellular reporter assay was implemented where the ligand binding domains of ROR α, β and γ were fused to the DNA binding domain of the transcriptional factor GAL4. RESULTS |
|
221 225 GAL4 protein In order to assess the in vivo selectivity profile of BIO399 a cellular reporter assay was implemented where the ligand binding domains of ROR α, β and γ were fused to the DNA binding domain of the transcriptional factor GAL4. RESULTS |
|
4 7 ROR protein_type The ROR-GAL4 fusion proteins were expressed in cells with the luciferase reporter gene under the control of a GAL4 promoter. RESULTS |
|
8 12 GAL4 protein The ROR-GAL4 fusion proteins were expressed in cells with the luciferase reporter gene under the control of a GAL4 promoter. RESULTS |
|
110 114 GAL4 protein The ROR-GAL4 fusion proteins were expressed in cells with the luciferase reporter gene under the control of a GAL4 promoter. RESULTS |
|
0 6 BIO399 chemical BIO399 inhibited the luciferase activity when added to the cells expressing the RORγ-GAL4 fusion with an in vivo IC50 of 42.5nM while showing >235 and 28 fold selectivity over cells expressing GAL4 fused to the LBD of ROR α or β, respectively (Table 1). RESULTS |
|
80 84 RORγ protein BIO399 inhibited the luciferase activity when added to the cells expressing the RORγ-GAL4 fusion with an in vivo IC50 of 42.5nM while showing >235 and 28 fold selectivity over cells expressing GAL4 fused to the LBD of ROR α or β, respectively (Table 1). RESULTS |
|
85 89 GAL4 protein BIO399 inhibited the luciferase activity when added to the cells expressing the RORγ-GAL4 fusion with an in vivo IC50 of 42.5nM while showing >235 and 28 fold selectivity over cells expressing GAL4 fused to the LBD of ROR α or β, respectively (Table 1). RESULTS |
|
113 117 IC50 evidence BIO399 inhibited the luciferase activity when added to the cells expressing the RORγ-GAL4 fusion with an in vivo IC50 of 42.5nM while showing >235 and 28 fold selectivity over cells expressing GAL4 fused to the LBD of ROR α or β, respectively (Table 1). RESULTS |
|
193 197 GAL4 protein BIO399 inhibited the luciferase activity when added to the cells expressing the RORγ-GAL4 fusion with an in vivo IC50 of 42.5nM while showing >235 and 28 fold selectivity over cells expressing GAL4 fused to the LBD of ROR α or β, respectively (Table 1). RESULTS |
|
211 214 LBD structure_element BIO399 inhibited the luciferase activity when added to the cells expressing the RORγ-GAL4 fusion with an in vivo IC50 of 42.5nM while showing >235 and 28 fold selectivity over cells expressing GAL4 fused to the LBD of ROR α or β, respectively (Table 1). RESULTS |
|
218 221 ROR protein_type BIO399 inhibited the luciferase activity when added to the cells expressing the RORγ-GAL4 fusion with an in vivo IC50 of 42.5nM while showing >235 and 28 fold selectivity over cells expressing GAL4 fused to the LBD of ROR α or β, respectively (Table 1). RESULTS |
|
222 223 α protein BIO399 inhibited the luciferase activity when added to the cells expressing the RORγ-GAL4 fusion with an in vivo IC50 of 42.5nM while showing >235 and 28 fold selectivity over cells expressing GAL4 fused to the LBD of ROR α or β, respectively (Table 1). RESULTS |
|
227 228 β protein BIO399 inhibited the luciferase activity when added to the cells expressing the RORγ-GAL4 fusion with an in vivo IC50 of 42.5nM while showing >235 and 28 fold selectivity over cells expressing GAL4 fused to the LBD of ROR α or β, respectively (Table 1). RESULTS |
|
4 7 LBS site The LBS of RORs share a high degree of similarity. RESULTS |
|
11 15 RORs protein_type The LBS of RORs share a high degree of similarity. RESULTS |
|
40 46 BIO399 chemical However, the inverse agonism trigger of BIO399, residue Met358, is a leucine in both RORα and β. RESULTS |
|
56 62 Met358 residue_name_number However, the inverse agonism trigger of BIO399, residue Met358, is a leucine in both RORα and β. RESULTS |
|
69 76 leucine residue_name However, the inverse agonism trigger of BIO399, residue Met358, is a leucine in both RORα and β. RESULTS |
|
85 89 RORα protein However, the inverse agonism trigger of BIO399, residue Met358, is a leucine in both RORα and β. RESULTS |
|
94 95 β protein However, the inverse agonism trigger of BIO399, residue Met358, is a leucine in both RORα and β. RESULTS |
|
29 35 BIO399 chemical This selectivity profile for BIO399 is attributed to the shorter leucine side chain in RORα and β which would not reach the phenylalanine on the AF2 helix further underscoring the role of Met358 as a trigger for RORγ specific inverse agonism (Fig. 7a). RESULTS |
|
65 72 leucine residue_name This selectivity profile for BIO399 is attributed to the shorter leucine side chain in RORα and β which would not reach the phenylalanine on the AF2 helix further underscoring the role of Met358 as a trigger for RORγ specific inverse agonism (Fig. 7a). RESULTS |
|
87 91 RORα protein This selectivity profile for BIO399 is attributed to the shorter leucine side chain in RORα and β which would not reach the phenylalanine on the AF2 helix further underscoring the role of Met358 as a trigger for RORγ specific inverse agonism (Fig. 7a). RESULTS |
|
96 97 β protein This selectivity profile for BIO399 is attributed to the shorter leucine side chain in RORα and β which would not reach the phenylalanine on the AF2 helix further underscoring the role of Met358 as a trigger for RORγ specific inverse agonism (Fig. 7a). RESULTS |
|
124 137 phenylalanine residue_name This selectivity profile for BIO399 is attributed to the shorter leucine side chain in RORα and β which would not reach the phenylalanine on the AF2 helix further underscoring the role of Met358 as a trigger for RORγ specific inverse agonism (Fig. 7a). RESULTS |
|
145 154 AF2 helix structure_element This selectivity profile for BIO399 is attributed to the shorter leucine side chain in RORα and β which would not reach the phenylalanine on the AF2 helix further underscoring the role of Met358 as a trigger for RORγ specific inverse agonism (Fig. 7a). RESULTS |
|
188 194 Met358 residue_name_number This selectivity profile for BIO399 is attributed to the shorter leucine side chain in RORα and β which would not reach the phenylalanine on the AF2 helix further underscoring the role of Met358 as a trigger for RORγ specific inverse agonism (Fig. 7a). RESULTS |
|
212 216 RORγ protein This selectivity profile for BIO399 is attributed to the shorter leucine side chain in RORα and β which would not reach the phenylalanine on the AF2 helix further underscoring the role of Met358 as a trigger for RORγ specific inverse agonism (Fig. 7a). RESULTS |
|
13 17 RORα protein Furthermore, RORα contains two phenylalanine residues in its LBS whereas RORβ and γ have a leucine in the same position (Fig. 6b). RESULTS |
|
31 44 phenylalanine residue_name Furthermore, RORα contains two phenylalanine residues in its LBS whereas RORβ and γ have a leucine in the same position (Fig. 6b). RESULTS |
|
61 64 LBS site Furthermore, RORα contains two phenylalanine residues in its LBS whereas RORβ and γ have a leucine in the same position (Fig. 6b). RESULTS |
|
73 77 RORβ protein Furthermore, RORα contains two phenylalanine residues in its LBS whereas RORβ and γ have a leucine in the same position (Fig. 6b). RESULTS |
|
82 83 γ protein Furthermore, RORα contains two phenylalanine residues in its LBS whereas RORβ and γ have a leucine in the same position (Fig. 6b). RESULTS |
|
91 98 leucine residue_name Furthermore, RORα contains two phenylalanine residues in its LBS whereas RORβ and γ have a leucine in the same position (Fig. 6b). RESULTS |
|
28 41 phenylalanine residue_name We hypothesize that the two phenylalanine residues in the LBS of RORα occlude the dihydrobenzoxazepinone ring system of BIO399 from binding it and responsible for the increase in selectivity for RORα over β. RESULTS |
|
58 61 LBS site We hypothesize that the two phenylalanine residues in the LBS of RORα occlude the dihydrobenzoxazepinone ring system of BIO399 from binding it and responsible for the increase in selectivity for RORα over β. RESULTS |
|
65 69 RORα protein We hypothesize that the two phenylalanine residues in the LBS of RORα occlude the dihydrobenzoxazepinone ring system of BIO399 from binding it and responsible for the increase in selectivity for RORα over β. RESULTS |
|
82 104 dihydrobenzoxazepinone chemical We hypothesize that the two phenylalanine residues in the LBS of RORα occlude the dihydrobenzoxazepinone ring system of BIO399 from binding it and responsible for the increase in selectivity for RORα over β. RESULTS |
|
120 126 BIO399 chemical We hypothesize that the two phenylalanine residues in the LBS of RORα occlude the dihydrobenzoxazepinone ring system of BIO399 from binding it and responsible for the increase in selectivity for RORα over β. RESULTS |
|
195 199 RORα protein We hypothesize that the two phenylalanine residues in the LBS of RORα occlude the dihydrobenzoxazepinone ring system of BIO399 from binding it and responsible for the increase in selectivity for RORα over β. RESULTS |
|
205 206 β protein We hypothesize that the two phenylalanine residues in the LBS of RORα occlude the dihydrobenzoxazepinone ring system of BIO399 from binding it and responsible for the increase in selectivity for RORα over β. RESULTS |
|
47 60 benzoxazinone chemical We have identified a novel series of synthetic benzoxazinone ligands which modulate the transcriptional activity of RORγ in a FRET based assay. CONCL |
|
116 120 RORγ protein We have identified a novel series of synthetic benzoxazinone ligands which modulate the transcriptional activity of RORγ in a FRET based assay. CONCL |
|
126 142 FRET based assay experimental_method We have identified a novel series of synthetic benzoxazinone ligands which modulate the transcriptional activity of RORγ in a FRET based assay. CONCL |
|
6 25 partial proteolysis experimental_method Using partial proteolysis we show a conformational change which destabilizes the AF2 helix of RORγ when the inverse agonist BIO399 binds. CONCL |
|
81 90 AF2 helix structure_element Using partial proteolysis we show a conformational change which destabilizes the AF2 helix of RORγ when the inverse agonist BIO399 binds. CONCL |
|
94 98 RORγ protein Using partial proteolysis we show a conformational change which destabilizes the AF2 helix of RORγ when the inverse agonist BIO399 binds. CONCL |
|
108 123 inverse agonist protein_state Using partial proteolysis we show a conformational change which destabilizes the AF2 helix of RORγ when the inverse agonist BIO399 binds. CONCL |
|
124 130 BIO399 chemical Using partial proteolysis we show a conformational change which destabilizes the AF2 helix of RORγ when the inverse agonist BIO399 binds. CONCL |
|
8 12 RORγ protein The two RORγ co-crystal structures reported here show how a small change to the core ring system can modulate the mode of action from agonist (BIO592) to inverse agonism (BIO399). CONCL |
|
13 34 co-crystal structures evidence The two RORγ co-crystal structures reported here show how a small change to the core ring system can modulate the mode of action from agonist (BIO592) to inverse agonism (BIO399). CONCL |
|
134 141 agonist protein_state The two RORγ co-crystal structures reported here show how a small change to the core ring system can modulate the mode of action from agonist (BIO592) to inverse agonism (BIO399). CONCL |
|
143 149 BIO592 chemical The two RORγ co-crystal structures reported here show how a small change to the core ring system can modulate the mode of action from agonist (BIO592) to inverse agonism (BIO399). CONCL |
|
171 177 BIO399 chemical The two RORγ co-crystal structures reported here show how a small change to the core ring system can modulate the mode of action from agonist (BIO592) to inverse agonism (BIO399). CONCL |
|
67 71 RORγ protein Finally, we are reporting a newly identified trigger for achieving RORγ specific inverse agonism in an in vivo setting through Met358 which perturbs the agonist conformation of the AF2 helix and prevents coactivator protein binding. CONCL |
|
127 133 Met358 residue_name_number Finally, we are reporting a newly identified trigger for achieving RORγ specific inverse agonism in an in vivo setting through Met358 which perturbs the agonist conformation of the AF2 helix and prevents coactivator protein binding. CONCL |
|
153 160 agonist protein_state Finally, we are reporting a newly identified trigger for achieving RORγ specific inverse agonism in an in vivo setting through Met358 which perturbs the agonist conformation of the AF2 helix and prevents coactivator protein binding. CONCL |
|
181 190 AF2 helix structure_element Finally, we are reporting a newly identified trigger for achieving RORγ specific inverse agonism in an in vivo setting through Met358 which perturbs the agonist conformation of the AF2 helix and prevents coactivator protein binding. CONCL |
|
|
