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

Arch Biochem Biophys. 2001 Jul 15;391(2):180-7. doi: 10.1006/abbi.2001.2415.

Abstract

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-6 (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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alanine / metabolism
  • Amino Acid Sequence
  • Amino Acid Substitution
  • Conserved Sequence
  • Cytochrome P-450 Enzyme System*
  • Hydroxylation
  • Leucine / metabolism
  • Mixed Function Oxygenases / genetics
  • Mixed Function Oxygenases / metabolism*
  • Models, Molecular
  • Molecular Sequence Data
  • Phenylalanine / genetics
  • Phenylalanine / metabolism
  • Rosales / enzymology*
  • Sequence Homology, Amino Acid
  • Substrate Specificity
  • Valine / metabolism

Substances

  • Phenylalanine
  • Cytochrome P-450 Enzyme System
  • Mixed Function Oxygenases
  • fatty acid alpha-hydroxylase
  • cytochrome P-450 CYP94A2 (Vicia sativa)
  • Leucine
  • Valine
  • Alanine