Design and preparation of the class B G protein-coupled receptors GLP-1R and GCGR for F-19-NMR studies in solution

doi:10.1111/febs.15686

	Design and preparation of the class B G protein-coupled receptors GLP-1R and GCGR for F-19-NMR studies in solution
	Wang, Huixia1,2,3,4 ; Hu, Wanhui1,6 ; Liu, Dongsheng1 ; Wuthrich, Kurt1,5
	2021-07
发表期刊	FEBS JOURNAL (IF:5.5[JCR-2023],5.5[5-Year])
ISSN	1742-464X
EISSN	1742-4658
卷号	288 期号:13 页码:4053-4063
发表状态	已发表
DOI	10.1111/febs.15686
摘要	The human glucagon-like peptide-1 receptor (GLP-1R) and the glucagon receptor (GCGR) are class B G protein-coupled receptors (GPCRs) that are activated by interactions with, respectively, the glucagon-like peptide-1 (GLP-1) and glucagon (GCG). These polypeptide hormones are involved in the regulation of lipid and cholic acid metabolism, and thus play an important role in the pathogenesis of glucose metabolism and diabetes mellitus, which attracts keen interest of these GPCRs as drug targets. GLP-1R and GCGR have therefore been extensively investigated by X-ray crystallography and cryo-electron microscopy (cryo-EM), so that their structures are well known. Here, we present the groundwork for using nuclear magnetic resonance (NMR) spectroscopy in solution to complement the molecular architectures with information on intramolecular dynamics and on the thermodynamics and kinetics of interactions with physiological ligands and extrinsic drug candidates. This includes the generation of novel, near-wild-type constructs of GLP-1R and GCGR, optimization of the solution conditions for NMR studies in detergent micelles and in nanodiscs, post-translational chemical introduction of fluorine-19 NMR probes, and sequence-specific assignments of the F-19-labels attached to indigenous cysteines. Addition of the negative allosteric modulator (NAM) NNC0640 was critically important for obtaining the long-time stability needed for our NMR experiments, and we report on novel insights into the allosteric effects arising from binding of NNC0640 to the transmembrane domain of GLP-1R (GLP-1R[TMD]).
关键词	F-19-NMR glucagon receptor glucagon-like peptide-1 receptor GPCR negative allosteric modulator
收录类别	SCI ; SCIE
语种	英语
资助项目	National Natural Science Foundation of China[31971153][31670733]
WOS研究方向	Biochemistry & Molecular Biology
WOS类目	Biochemistry & Molecular Biology
WOS记录号	WOS:000605998500001
出版者	WILEY
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/125037
专题	生命科学与技术学院_博士生 iHuman研究所_特聘教授组_Kurt Wuthrich组
通讯作者	Wuthrich, Kurt
作者单位	1.ShanghaiTech Univ, iHuman Inst, Shanghai 201210, Peoples R China 2.Univ Chinese Acad Sci, Beijing, Peoples R China 3.ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai, Peoples R China 4.Chinese Acad Sci, Shanghai Inst Biochem & Cell Biol, CAS Ctr Excellence Mol Cell Sci, Shanghai, Peoples R China 5.Scripps Res, Dept Integrat Struct & Computat Biol, La Jolla, CA USA 6.Shanghai Henlius Biotech Inc, Shanghai, Peoples R China
第一作者单位	iHuman研究所; 生命科学与技术学院
通讯作者单位	iHuman研究所
第一作者的第一单位	iHuman研究所
推荐引用方式 GB/T 7714	Wang, Huixia,Hu, Wanhui,Liu, Dongsheng,et al. Design and preparation of the class B G protein-coupled receptors GLP-1R and GCGR for F-19-NMR studies in solution[J]. FEBS JOURNAL,2021,288(13):4053-4063.
APA	Wang, Huixia,Hu, Wanhui,Liu, Dongsheng,&Wuthrich, Kurt.(2021).Design and preparation of the class B G protein-coupled receptors GLP-1R and GCGR for F-19-NMR studies in solution.FEBS JOURNAL,288(13),4053-4063.
MLA	Wang, Huixia,et al."Design and preparation of the class B G protein-coupled receptors GLP-1R and GCGR for F-19-NMR studies in solution".FEBS JOURNAL 288.13(2021):4053-4063.