6KK7

Structure of thermal-stabilised(M6) human GLP-1 receptor transmembrane domain

Summary for 6KK7

• Entry DOI: 10.2210/pdb6kk7/pdb
• Related: 5VEW
• Descriptor: Glucagon-like peptide 1 receptor,Endolysin,Glucagon-like peptide 1 receptor, N-{4-[(R)-(3,3-dimethylcyclobutyl)({6-[4-(trifluoromethyl)-1H-imidazol-1-yl]pyridin-3-yl}amino)methyl]benzene-1-carbonyl}-beta-alanine (2 entities in total)
• Functional Keywords: gpcr, seven-transmembrane, allosteric modulators, diabetes, signaling protein
• Biological source: Homo sapiens (Human); More
• Total number of polymer chains: 2
• Total formula weight: 106129.51

Authors

Song, G. (deposition date: 2019-07-23, release date: 2019-11-13, Last modification date: 2024-10-16)

Primary citation

Xu, Y.,Wang, Y.,Wang, Y.,Liu, K.,Peng, Y.,Yao, D.,Tao, H.,Liu, H.,Song, G.
Mutagenesis facilitated crystallization of GLP-1R.
Iucrj, 6:996-1006, 2019

PubMed Abstract: The class B family of G-protein-coupled receptors (GPCRs) has long been a paradigm for peptide hormone recognition and signal transduction. One class B GPCR, the glucagon-like peptide-1 receptor (GLP-1R), has been considered as an anti-diabetes drug target and there are several peptidic drugs available for the treatment of this overwhelming disease. The previously determined structures of inactive GLP-1R in complex with two negative allosteric modulators include ten thermal-stabilizing mutations that were selected from a total of 98 designed mutations. Here we systematically summarize all 98 mutations we have tested and the results suggest that the mutagenesis strategy that strengthens inter-helical hydro-phobic interactions shows the highest success rate. We further investigate four back mutations by thermal-shift assay, crystallization and molecular dynamic simulations, and conclude that mutation I196F increases thermal stability intrinsically and that mutation S271A decreases crystal packing entropy extrinsically, while mutations S193C and M233C may be dispensable since these two cysteines are not di-sulfide-linked. Our results indicate intrinsic connections between different regions of GPCR transmembrane helices and the current data suggest a general mutagenesis principle for structural determination of GPCRs and other membrane proteins.

PubMed: 31709055
DOI: 10.1107/S2052252519013496

Experimental method

X-RAY DIFFRACTION (3.1 Å)