Mutagenesis facilitated crystallization of GLP-1R

doi:10.1107/S2052252519013496

	Mutagenesis facilitated crystallization of GLP-1R
	Xu, Yueming 1,2; Wang, Yuxia3 ; Wang, Yang 4; Liu, Kaiwen3,5 ; Peng, Yao3 ; Yao, Deqiang3 ; Tao, Houchao3 ; Liu, Haiguang 4; Song, Gaojie 1,2
	2019-11
发表期刊	IUCRJ (IF:2.9[JCR-2023],3.3[5-Year])
ISSN	2052-2525
卷号	6 页码:996-1006
发表状态	已发表
DOI	10.1107/S2052252519013496
摘要	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 hydrophobic 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 I196(2.66b)F increases thermal stability intrinsically and that mutation S271(4.47b)A decreases crystal packing entropy extrinsically, while mutations S193(2.63b)C and M233(3.36b)C may be dispensable since these two cysteines are not disulfide-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.
关键词	mutations G-protein-coupled receptors glucagon-like peptide-1 receptor membrane proteins molecular dynamic simulations crystallization
收录类别	SCI ; SCIE
语种	英语
资助项目	National Key R&D Program of China[2018YFA0507001]
WOS研究方向	Chemistry ; Crystallography ; Materials Science
WOS类目	Chemistry, Multidisciplinary ; Crystallography ; Materials Science, Multidisciplinary
WOS记录号	WOS:000495396200004
出版者	INT UNION CRYSTALLOGRAPHY
WOS关键词	GLUCAGON-LIKE PEPTIDE-1 ; CRYO-EM STRUCTURE ; CRYSTAL-STRUCTURE ; RECEPTOR ; MECHANISMS ; PROTEINS ; DATABASE ; COMPLEX ; BINDING
原始文献类型	Article
引用统计	正在获取...
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/102237
专题	生命科学与技术学院_博士生 iHuman研究所_特聘教授组_Raymond Stevens组 iHuman研究所_PI研究组_刘志杰组 iHuman研究所_PI研究组_陶厚朝组 iHuman研究所_科学装置(X)_膜蛋白同步辐射线站
通讯作者	Song, Gaojie
作者单位	1.East China Normal Univ, Inst Biomed Sci, Shanghai Key Lab Regulatory Biol, Shanghai 200241, Peoples R China 2.East China Normal Univ, Sch Life Sci, Shanghai 200241, Peoples R China 3.ShanghaiTech Univ, iHuman Inst, 393 Middle Huaxia Rd, Shanghai 201210, Peoples R China 4.Beijing Computat Sci Res Ctr, Complex Syst Div, Beijing 100193, Peoples R China 5.ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai 201210, Peoples R China
推荐引用方式 GB/T 7714	Xu, Yueming,Wang, Yuxia,Wang, Yang,et al. Mutagenesis facilitated crystallization of GLP-1R[J]. IUCRJ,2019,6:996-1006.
APA	Xu, Yueming.,Wang, Yuxia.,Wang, Yang.,Liu, Kaiwen.,Peng, Yao.,...&Song, Gaojie.(2019).Mutagenesis facilitated crystallization of GLP-1R.IUCRJ,6,996-1006.
MLA	Xu, Yueming,et al."Mutagenesis facilitated crystallization of GLP-1R".IUCRJ 6(2019):996-1006.