Distal mutation V486M disrupts the catalytic activity of DPP4 by affecting the flap of the propeller domain  

作　　者：Teng-teng Li Cheng Peng Ji-qiu Wang Zhi-jian Xu Ming-bo Su Jia Li Wei-liang Zhu Jing-ya Li 

机构地区：[1]State Key Laboratory of Drug Research,the National Drug Screening Center,Shanghai Institute of Materia Medica,Chinese Academy of Sciences,Shanghai,201203,China [2]School of Life Science and Technology,ShanghaiTech University,Shanghai,201210,China [3]CAS Key Laboratory of Receptor Research,Drug Discovery and Design Center,Shanghai Institute of Materia Medica,Chinese Academy of Sciences,Shanghai,201203,China [4]School of Pharmacy,University of Chinese Academy of Sciences,Beijing,100049,China [5]Department of Endocrinology and Metabolism,China National Research Center for Metabolic Diseases,National Key Laboratory for Medical Genomes,Ruijin Hospital,Shanghai Jiao Tong University School of Medicine(SJTUSM),Shanghai,200025,China

出　　处：《Acta Pharmacologica Sinica》2022年第8期2147-2155,共9页中国药理学报（英文版）

基　　金：supported by the National Natural Science Foundation of China(No.92057116);the National Science and Technology Major Project(2018ZX09711002-018);the Strategic Priority Research Program of Chinese Academy of Sciences grant(XDA12040204);the Shanghai Commission of Science and Technology(18431900900);National Key R&D Program of China(2016YFA0502301 and 2017YFB0202601).

摘　　要：Dipeptidyl peptidase-4(DPP4)plays a crucial role in regulating the bioactivity of glucagon-like peptide-1(GLP-1)that enhances insulin secretion and pancreaticβ-cell proliferation,making it a therapeutic target for type 2 diabetes.Although the crystal structure of DPP4 has been determined,its structure-function mechanism is largely unknown.Here,we examined the biochemical properties of sporadic human DPP4 mutations distal from its catalytic site,among which V486M ablates DPP4 dimerization and causes loss of enzymatic activity.Unbiased molecular dynamics simulations revealed that the distal V486M mutation induces a local conformational collapse in aβ-propeller loop(residues 234–260,defined as the flap)and disrupts the dimerization of DPP4.The“open/closed”conformational transitions of the flap whereby capping the active site,are involved in the enzymatic activity of DPP4.Further site-directed mutagenesis guided by theoretical predictions verified the importance of the conformational dynamics of the flap for the enzymatic activity of DPP4.Therefore,the current studies that combined theoretical modeling and experimental identification,provide important insights into the biological function of DPP4 and allow for the evaluation of directed DPP4 genetic mutations before initiating clinical applications and drug development.

关 键 词：DPP4 distal mutation enzymatic activity molecular dynamics simulation structure-function mechanism 

分 类 号：R285.5[医药卫生—中药学]