Journal: bioRxiv

Article Title: Structural Basis of GPCR-G Protein Pre-coupling and Activation: Insights from CCR1-Gi Complex

doi: 10.1101/2024.11.01.621549

Figure Lengend Snippet: a , Comparison of the interface between the α5 helix of Gαi and the cytoplasmic region of CCR1 in the active (green) and pre-coupled (dodger blue) states. The alignment is performed using the receptor as the reference. b-c, Agonist-induced changes in luciferase signal measured by NanoBiT between CCR1-SmBiT and Gα-LgBiT ( b ), and between Gγ-SmBiT and Gα-LgBiT ( c ). Bars and error bars indicate the mean and SEM, respectively. N = 6 ( b ) and 4 ( c ) independent experiments, performed with single replicates. d, Orthogonal views of the structures of the CCR1-Gi heterotrimer complex in the active and pre-coupled states, colored by subunit. CCR1 is shown in green (active state) and dodger blue (pre-coupled state), Gαi1 in gold (active state) and yellow (pre-coupled state), Gβ in rosy brown, Gγ in light blue, and CCL15 in brown. e-f, Luciferase complementation between various GPCRs (CCR1, CCR2, CCR4, CCR5, CXCR1, CXCR2, CX3CR1, and 5HT 1 R) fused to SmBiT and Gα-LgBiT decreases in response to agonist ( e ), whereas luciferase complementation between other GPCRs (NPY2R, FPR1, FPR2, CB1, AT 1 R, and AT 2 R) fused to SmBiT and Gα-LgBiT increases in response to agonist ( f ). N = 6-8 independent experiments, performed with single replicates. g, The model of GPCR activation in which the C-terminal residues of the Gαi α5 helix (red), while disordered in the pre-coupled state (blue), play a crucial role in the transition to the fully active state (green) and subsequent G protein dissociation

Article Snippet: Angiotensin II (HY-13948) for AT1R and AT2R, Serotonin(HY-B1473) for 5HT1eR, NPY protein(HY-P71063) for NPY2R,CX3CL1 protein(HY-P7180) for CX3CR1 are from MedChemexpress.

Techniques: Comparison, Luciferase, Activation Assay