Structural Snapshots for the Conformation-dependent Catalysis by Human Medium-chain Acyl-coenzyme A Synthetase ACSM2A

被引：90

作者：

Kochan, Grazyna ^{[1
]}

Pilka, Ewa S. ^{[1
]}

von Delft, Frank ^{[1
]}

Oppermann, Udo ^{[1
,2
]}

Yue, Wyatt W. ^{[1
]}

机构：

[1] Struct Genom Consortium, Oxford OX3 7DU, England

[2] Botnar Res Ctr, Oxford Biomed Res, Oxford OX3 7LD, England

来源：

JOURNAL OF MOLECULAR BIOLOGY | 2009年 / 388卷 / 05期

基金：

英国惠康基金;

关键词：

fatty acid; coenzyme A; adenylation; thioesterification; synthetase; NONRIBOSOMAL PEPTIDE SYNTHETASE; FORMING ENZYME SUPERFAMILY; CRYSTAL-STRUCTURE; SUBSTRATE-BINDING; MECHANISM; PURIFICATION; FRACTION; RESIDUE; SITE; DLTA;

D O I：

10.1016/j.jmb.2009.03.064

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Acyl-CoA synthetases belong to the superfamily of adenylate-forming enzymes, and catalyze the two-step activation of fatty acids or carboxylate-containing xenobiotics. The carboxylate Substrate first reacts with ATP to form an acyl-adenylate intermediate, which then reacts with CoA to produce in acyl-CoA ester. Here, we report the first crystal structure of a medium-chain acyl-CoA synthetase ACSM2A, in a series Of substrate/product/cofactor complexes central to the catalytic mechanism. We observed a substantial rearrangement between the N- and C-terminal domains, driven purely by the identity of the bound ligand in the active site. Our structures allowed LIS to identify the presence or absence of the ATP pyrophosphates as the conformational switch, and elucidated new mechanistic details, including the role of invariant Lys557 and a divalent magnesium ion in coordinating the ATP pyrophosphates, as well as the involvement of a Gly-rich P-loop and the conserved Arg472-Glu365 salt bridge in the domain rearrangement. (C) 2009 Elsevier Ltd. All rights reserved.

引用

页码：997 / 1008

页数：12

共 47 条

[11] Structure of the EntB multidomain nonribosomal peptide synthetase and functional analysis of its interaction with the EntE adenylation domain [J].