A conservative amino acid substitution alters the regiospecificity of CYP94A2, a fatty acid hydroxylase from the plant Vicia sativa

被引:35
作者
Kahn, RA [1 ]
Le Bouquin, R [1 ]
Pinot, F [1 ]
Benveniste, I [1 ]
Durst, F [1 ]
机构
[1] Inst Biol Mol Plantes, CNRS UPR 406, Dept Enzymol Cellulaire & Mol, F-67083 Strasbourg, France
关键词
cytochrome P450; CYP94A2; Vicia sativa; fatty acid hydroxylase; omega-hydroxylation; (omega-1)-hydroxylation; site-directed mutagenesis; SRS-6; Phe494; regioselectivity; active site;
D O I
10.1006/abbi.2001.2415
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
Fatty acid omega -hydroxylation is involved in the biosynthesis of the plant cuticle, formation of plant defense signaling molecules, and possibly in the rapid catabolism of free fatty acids liberated under stress conditions. CYP94A2 is a cytochrome P450-dependent medium-chain fatty acid hydroxylase that was recently isolated from Vicia sativa, Contrary to CYP94A1 and CYP86A1, two other fatty acid hydroxylases previously characterized in V. sativa and Arabidopsis thaliana, CYP94A2 is not a strict omega -hydroxylase, but exhibits chain-length-dependent regioselectivity of oxidative attack. Sequence alignments of CYP94A2 with CYP94A1 and molecular modeling studies suggested that F494, located in SRS-B (substrate recognition site) was involved in substrate recognition and positioning. Indeed, a conservative amino acid substitution at that position markedly altered the regiospecificity of CYP94A2. The observed shift from omega toward omega -1 hydroxylation was prominent with lauric acid as substrate and declined with increasing fatty acid chain length. (C) 2001 Academic Press.
引用
收藏
页码:180 / 187
页数:8
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