期刊
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS
卷 520, 期 2, 页码 99-107出版社
ELSEVIER SCIENCE INC
DOI: 10.1016/j.abb.2012.02.010
关键词
Enterococcus faecalis; Azoreductase; Azo dye; NADH; Site-directed mutagenesis
资金
- National Center for Toxicological Research, United States Food and Drug Administration
- Oak Ridge Institute for Science and Education
AzoA from Enterococcus faecalis is a member of the polymeric Flavin-dependent NADH-preferred azoreductase group. Little is known about the binding and interaction of NADH and azo dye in the azoreductase group. A synergetic strategy based on computational prediction, reverse genetics validation coupled with site-directed mutagenesis, and reconstruction of mutation network was used to investigate the binding and interaction of NADH and a model azo dye, Methyl Red, with AzoA. Methyl Red and NADH interacted in a unique binding mode in which the benzoic acid moiety of Methyl Red and the nicotinamide ring of NADH were not parallel to the Flavin isoalloxazine ring, but lay against it at angles of similar to 45 degrees and similar to 35 degrees, respectively. The adenine ribose moiety of NADH was surrounded by loop l2 on chain B and alpha 3 on chain A in a typical Rossmann fold. There were 12 and 19 amino acid residues that could participate in the binding of Methyl Red and NADH, respectively, especially the residues Tyr-129 and Asp-184. The functional perturbation effects of 13 residues, including Tyr-129 and Asp-184, were mapped to reconstruct the mutation network, which confirmed the proposed binding modes and also provided insights into the interaction among NADH, FMN and Methyl Red. Published by Elsevier Inc.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据