Journal: Nature Structural & Molecular Biology

Article Title: Structural insights into the activation and inhibition of CXC chemokine receptor 3

doi: 10.1038/s41594-023-01175-5

Figure Lengend Snippet: a – c , Density maps and cartoon models of CXCR3–CXCL11–DNG i -scFv16 ( a ), CXCR3–PS372424–DNG i -scFv16 ( b ) and CXCR3–VUF11222–DNG i -scFv16 ( c ). Receptor, ligand, DNG αi , G β , G γ and scFv16 are colored violet, cyan, blue, green, orange and gray, respectively. PS372424 and VUF11222 are shown as spheres. d , Density map and cartoon model of CXCR3 κOR –SCH546738-Nb6. CXCR3, κOR fragment, SCH546738 and Nb6 are colored violet, orange, cyan and gray, respectively. SCH546738 is shown with spheres.

Article Snippet: Fig. 6 Discovery of an allosteric site in the structure of CXCR3 occupied by SCH546738. a , The chemical structure of SCH546738. b , Molecular dynamics simulation of SCH546738 binding.

Techniques:

Journal: Nature Structural & Molecular Biology

Article Title: Structural insights into the activation and inhibition of CXC chemokine receptor 3

doi: 10.1038/s41594-023-01175-5

Figure Lengend Snippet: a Elution curve of CXCR3-CXCL11-DNG i -scFv16 complex on Superose 6 Increase column. b SDS-PAGE of CXCR3-CXCL11-DNG i -scFv16 complex after size-exclusion chromatography. c Cryo-EM image of CXCR3-CXCL11-DNG i -scFv16 particles (a representative of 5,500 micrographs). d Elution curve of CXCR3-PS372424-DNG i -scFv16 complex on Superose 6 Increase column. e SDS-PAGE of CXCR3-PS372424-DNG i -scFv16 complex after size-exclusion chromatography. f Cryo-EM image of CXCR3-PS372424-DNG i -scFv16 particles (a representative of 3,185 micrographs). g Elution curve of CXCR3-VUF11222-DNG i -scFv16 complex on Superose 6 Increase column. h SDS-PAGE of CXCR3-VUF11222-DNG i -scFv16 complex after size-exclusion chromatography. i Cryo-EM image of CXCR3-VUF11222-DNG i -scFv16 particles (a representative of 2,891 micrographs). j Elution curve of CXCR3 κOR -SCH546738-Nb6 complex on Superose 6 Increase column. k SDS-PAGE of CXCR3 κOR -SCH546738-Nb6 complex after size-exclusion chromatography. l Cryo-EM image of CXCR3 κOR -SCH546738-Nb6 particles (a representative of 12,944 micrographs). In b, e, h, and k, data shown are representative of two independent experiments.

Article Snippet: Fig. 6 Discovery of an allosteric site in the structure of CXCR3 occupied by SCH546738. a , The chemical structure of SCH546738. b , Molecular dynamics simulation of SCH546738 binding.

Techniques: SDS Page, Size-exclusion Chromatography, Cryo-EM Sample Prep

Journal: Nature Structural & Molecular Biology

Article Title: Structural insights into the activation and inhibition of CXC chemokine receptor 3

doi: 10.1038/s41594-023-01175-5

Figure Lengend Snippet: a Cryo-EM data processing of CXCR3-CXCL11-DNG i -scFv16. The local resolution map and the FSC curve are presented as well. b Cryo-EM data processing of CXCR3-PS372424-DNG i -scFv16. The local resolution map and the FSC curve are presented as well. c Cryo-EM data processing of CXCR3-VUF11222-DNG i -scFv16. The local resolution map and the FSC curve are presented as well. d Cryo-EM data processing of CXCR3 κOR -SCH546738-Nb6. The local resolution map and the FSC curve are presented as well.

Article Snippet: Fig. 6 Discovery of an allosteric site in the structure of CXCR3 occupied by SCH546738. a , The chemical structure of SCH546738. b , Molecular dynamics simulation of SCH546738 binding.

Techniques: Cryo-EM Sample Prep

Journal: Nature Structural & Molecular Biology

Article Title: Structural insights into the activation and inhibition of CXC chemokine receptor 3

doi: 10.1038/s41594-023-01175-5

Figure Lengend Snippet: Statistics of data collection, data processing, model refinement and validation

Article Snippet: Fig. 6 Discovery of an allosteric site in the structure of CXCR3 occupied by SCH546738. a , The chemical structure of SCH546738. b , Molecular dynamics simulation of SCH546738 binding.

Techniques: Biomarker Discovery

Journal: Nature Structural & Molecular Biology

Article Title: Structural insights into the activation and inhibition of CXC chemokine receptor 3

doi: 10.1038/s41594-023-01175-5

Figure Lengend Snippet: a Density maps of the transmembrane helixes and CXCL11 in the structure of CXCR3-CXCL11-DNG i -scFv16. b Density maps of the transmembrane helixes and PS372424 in the structure of CXCR3- PS372424 -DNG i -scFv16. c Density maps of the transmembrane helixes and VUF11222 in the structure of CXCR3- VUF11222 -DNG i -scFv16. d Density maps of the transmembrane helixes and SCH546738 in the structure of CXCR3 κOR - SCH546738-Nb6.

Article Snippet: Fig. 6 Discovery of an allosteric site in the structure of CXCR3 occupied by SCH546738. a , The chemical structure of SCH546738. b , Molecular dynamics simulation of SCH546738 binding.

Techniques:

Journal: Nature Structural & Molecular Biology

Article Title: Structural insights into the activation and inhibition of CXC chemokine receptor 3

doi: 10.1038/s41594-023-01175-5

Figure Lengend Snippet: a , Comparison of the insertion depth of the three agonists. The receptors are shown as cartoons and the ligands are shown as sticks. CXCL11, PS372424 and VUF11222 are colored violet, yellow and blue, respectively. b , Superposition of CXCL11-activated CXCR3 (violet) and PS372424-activated CXCR3 (yellow). c , Superposition of CXCL11-activated CXCR3 (violet) and VUF11222-activated CXCR3 (blue). d , The residues in CXCR3 involved in the interactions with CXCL11 (violet), PS372424 (yellow) and VUF11222 (blue). The residues are shown as sticks, and the CXCL11-activated CXCR3 are shown as cartoons. e , Comparison of CXCL11-activated CXCR3 (violet) and SCH546378-inhibited CXCR3 (green). f , The packing between TM3 and TM6 in the CXCR3 inhibited by antagonist SCH546738. CXCR3 is shown as a cartoon model and colored green and residues involved in TM packing are shown as sticks. g , The packing between TM5 and TM6 in the CXCR3 activated by chemokine CXCL11. CXCR3 is shown as a cartoon model and colored violet, and residues involved in TM packing are shown as sticks. h , Outward displacement of TM6 on CXCL11 binding leads to the exposure of the G αi binding pocket. The CXCL11-activated CXCR3, SCH546378-inhibited CXCR3 and G αi are shown as cartoon and colored violet, green and gray, respectively.

Article Snippet: Fig. 6 Discovery of an allosteric site in the structure of CXCR3 occupied by SCH546738. a , The chemical structure of SCH546738. b , Molecular dynamics simulation of SCH546738 binding.

Techniques: Comparison, Binding Assay

Journal: Nature Structural & Molecular Biology

Article Title: Structural insights into the activation and inhibition of CXC chemokine receptor 3

doi: 10.1038/s41594-023-01175-5

Figure Lengend Snippet: a , The chemical structure of SCH546738. b , Molecular dynamics simulation of SCH546738 binding. Shown are the cryo-EM structure and the top three cluster centroids. The size of each cluster is represented as the percentage of the total number of frames in the trajectories. c , A general view of the SCH546738 binding pocket in CXCR3. SCH546738 and CXCR3 are shown as a stick model (colored gray) and surface model (colored by electronic potential), respectively. d , Interactions between SCH546738 (gray) and CXCR3 (green). SCH546738 and the residues involved in interactions are shown as sticks. e , cAMP responses of CXCR3 to CXCL11 in the presence of SCH546738 at different concentrations. f , cAMP responses of CXCR3 V261F to CXCL11 in the presence of SCH546738 at different concentrations. g , cAMP responses of CXCR3 A265F to CXCL11 in the presence of SCH546738 at different concentrations. In e – g , the data represent means ± s.e.m. ( n = 6 independent experiments). h , Comparison of the allosteric binding sites outside the TMs in class A GPCR. The receptors are shown as cartoons and the antagonists are shown as spheres. Allosteric binding sites are indicated by red circles.

Article Snippet: Fig. 6 Discovery of an allosteric site in the structure of CXCR3 occupied by SCH546738. a , The chemical structure of SCH546738. b , Molecular dynamics simulation of SCH546738 binding.

Techniques: Binding Assay, Cryo-EM Sample Prep, Comparison

Journal: Nature Structural & Molecular Biology

Article Title: Structural insights into the activation and inhibition of CXC chemokine receptor 3

doi: 10.1038/s41594-023-01175-5

Figure Lengend Snippet: The potencies of CXCL11 with the receptor or its mutants in the present of different concentrations of SCH546738

Article Snippet: Fig. 6 Discovery of an allosteric site in the structure of CXCR3 occupied by SCH546738. a , The chemical structure of SCH546738. b , Molecular dynamics simulation of SCH546738 binding.

Techniques: