Catalytic mechanism of Escherichia coli isopentenyl diphosphate isomerase involves Cys-67, Glu-116, and Tyr-104 as suggested by crystal structures of complexes with transition state analogues and irreversible inhibitors

J Biol Chem. 2003 Apr 4;278(14):11903-8. doi: 10.1074/jbc.M212823200. Epub 2003 Jan 22.

Abstract

Isopentenyl diphosphate (IPP):dimethylallyl diphosphate (DMAPP) isomerase is a key enzyme in the biosynthesis of isoprenoids. The reaction involves protonation and deprotonation of the isoprenoid unit and proceeds through a carbocationic transition state. Analysis of the crystal structures (2 A) of complexes of Escherichia coli IPP.DMAPPs isomerase with a transition state analogue (N,N-dimethyl-2-amino-1-ethyl diphosphate) and a covalently attached irreversible inhibitor (3,4-epoxy-3-methyl-1-butyl diphosphate) indicates that Glu-116, Tyr-104, and Cys-67 are involved in the antarafacial addition/elimination of protons during isomerization. This work provides a new perspective about the mechanism of the reaction.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Carbon-Carbon Double Bond Isomerases / antagonists & inhibitors
  • Carbon-Carbon Double Bond Isomerases / chemistry*
  • Carbon-Carbon Double Bond Isomerases / metabolism*
  • Catalytic Domain
  • Crystallography
  • Cysteine / chemistry
  • Epoxy Compounds / pharmacology
  • Escherichia coli / enzymology*
  • Glutamic Acid / chemistry
  • Hemiterpenes
  • Kinetics
  • Organophosphorus Compounds / pharmacology
  • Protein Structure, Secondary
  • Tyrosine / chemistry

Substances

  • Epoxy Compounds
  • Hemiterpenes
  • Organophosphorus Compounds
  • 3-methyl-3,4-epoxybutyl diphosphate
  • Glutamic Acid
  • Tyrosine
  • Carbon-Carbon Double Bond Isomerases
  • isopentenyldiphosphate delta-isomerase
  • Cysteine

Associated data

  • PDB/1NFS
  • PDB/1NFZ