生成顺式烯烃所需的模块化聚酮合酶酮还原酶结构域的结构和立体化学分析。
Structural and stereochemical analysis of a modular polyketide synthase ketoreductase domain required for the generation of a cis-alkene.
作者信息
Bonnett Shilah A, Whicher Jonathan R, Papireddy Kancharla, Florova Galina, Smith Janet L, Reynolds Kevin A
机构信息
Department of Chemistry, Portland State University, Portland, OR. 97201.
Chemical Biology Graduate Program, University of Michigan, Ann Arbor, Michigan 48109.
出版信息
Chem Biol. 2013 Jun 20;20(6):772-83. doi: 10.1016/j.chembiol.2013.04.014.
Abstract
The formation of an activated cis-3-cyclohexylpropenoic acid by Plm1, the first extension module of the phoslactomycin polyketide synthase, is proposed to occur through an L-3-hydroxyacyl-intermediate as a result of ketoreduction by an A-type ketoreductase (KR). Here, we demonstrate that the KR domain of Plm1 (PlmKR1) catalyzes the formation of an L-3-hydroxyacyl product. The crystal structure of PlmKR1 revealed a well-ordered active site with a nearby Trp residue characteristic of A-type KRs. Structural comparison of PlmKR1 with B-type KRs that produce D-3-hydroxyacyl intermediates revealed significant differences. The active site of cofactor-bound A-type KRs is in a catalysis-ready state, whereas cofactor-bound B-type KRs are in a precatalytic state. Furthermore, the closed lid loop in substrate-bound A-type KRs restricts active site access from all but one direction, which is proposed to control the stereochemistry of ketoreduction.
摘要
磷霉素聚酮合酶的首个延伸模块Plm1催化生成顺式-3-环己基丙烯酸，据推测这是通过一种A型酮还原酶（KR）进行酮还原，以L-3-羟基酰基中间体为媒介而发生的。在此，我们证明Plm1的KR结构域（PlmKR1）催化生成L-3-羟基酰基产物。PlmKR1的晶体结构显示其活性位点排列有序，附近有一个A型KR特有的色氨酸残基。将PlmKR1与生成D-3-羟基酰基中间体的B型KR进行结构比较，发现了显著差异。结合辅因子的A型KR的活性位点处于催化就绪状态，而结合辅因子的B型KR处于预催化状态。此外，底物结合型A型KR中闭合的盖子环限制了除一个方向外从其他所有方向进入活性位点，这被认为可控制酮还原的立体化学。
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