文献类型：期刊文献

英文题名：Structural insights into substrate and coenzyme preference by SDR family protein Gox2253 from Gluconobater oxydans

作者：Yin, Bo[1,2];Cui, Dongbing[1];Zhang, Lujia[1];Jiang, Shuiqin[1];Machida, Satoru[2];Yuan, Y. Adam[2,3,4];Wei, Dongzhi[1]

机构：[1]E China Univ Sci & Technol, New World Inst Biotechnol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Natl Univ Singapore, Dept Biol Sci, Singapore 117543, Singapore;[3]Natl Univ Singapore, Ctr Bioimaging Sci, Singapore 117543, Singapore;[4]Natl Univ Singapore Suzhou, Res Inst, Suzhou 215123, Jiangsu, Peoples R China

年份：2014

卷号：82

期号：11

起止页码：2925

外文期刊名：PROTEINS-STRUCTURE FUNCTION AND BIOINFORMATICS

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000344378300006)】；

基金：Grant sponsor: National University of Singapore (Suzhou) Research Institute; Grant number: R-2012-N-007; Grant sponsor: Tier 1 research fund from Singapore Ministry of Education; Grant number: R-154-000-618-112 (Y.A.Y.); Grant sponsor: The Fundamental Research Funds for the Central Universities, "973" Program; Grant number: 2012CB721003; Grant sponsor: National Natural Science Foundation of China; Grant number: 21276084/B060804 (D. W.). Bo Yin and Dongbing Cui contributed equally to this work.

语种：英文

外文关键词：crystal structure; short-chain dehydrogenases; reductases; G; oxydans Gox2253; substrate selection; coenzyme preference

摘要：Gox2253 from Gluconobacter oxydans belongs to the short-chain dehydrogenases/reductases family, and catalyzes the reduction of heptanal, octanal, nonanal, and decanal with NADPH. To develop a robust working platform to engineer novel G. oxydans oxidoreductases with designed coenzyme preference, we adopted a structure based rational design strategy using computational predictions that considers the number of hydrogen bonds formed between enzyme and docked coenzyme. We report the crystal structure of Gox2253 at 2.6 angstrom resolution, ternary models of Gox2253 mutants in complex with NADH/short-chain aldehydes, and propose a structural mechanism of substrate selection. Molecular dynamics simulation shows that hydrogen bonds could form between 2-hydroxyl group in the adenosine moiety of NADH and the side chain of Gox2253 mutant after arginine at position 42 is replaced with tyrosine or lysine. Consistent with the molecular dynamics prediction, Gox2253-R42Y/K mutants can use both NADH and NADPH as a coenzyme. Hence, the strategies here could provide a practical platform to engineer coenzyme selectivity for any given oxidoreductase and could serve as an additional consideration to engineer substrate-binding pockets. Proteins 2014; 82:2925-2935. (c) 2014 Wiley Periodicals, Inc.