X-ray crystallography reveals a reduced substrate complex of UDP-galactopyranose mutase poised for covalent catalysis by flavin

Biochemistry. 2009 Oct 6;48(39):9171-3. doi: 10.1021/bi901437v.

Abstract

The flavoenzyme uridine 5'-diphosphate galactopyranose mutase (UGM or Glf) catalyzes the interconversion of UDP-galactopyranose and UDP-galactofuranose. The latter is a key building block for cell wall construction in numerous pathogens, including Mycobacterium tuberculosis. Mechanistic studies of UGM suggested a novel role for the flavin, and we previously provided evidence that the catalytic mechanism proceeds through a covalent flavin-galactose iminium. Here, we describe 2.3 and 2.5 A resolution X-ray crystal structures of the substrate-bound enzyme in oxidized and reduced forms, respectively. In the latter, C1 of the substrate is 3.6 A from the nucleophilic flavin N5 position. This orientation is consistent with covalent catalysis by flavin.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Catalysis
  • Crystallography, X-Ray
  • Flavin-Adenine Dinucleotide / chemistry
  • Flavin-Adenine Dinucleotide / metabolism*
  • Intramolecular Transferases / chemistry*
  • Intramolecular Transferases / metabolism*
  • Klebsiella pneumoniae / enzymology
  • Oxidation-Reduction
  • Substrate Specificity

Substances

  • Flavin-Adenine Dinucleotide
  • Intramolecular Transferases
  • UDP-galactopyranose mutase

Associated data

  • PDB/3INR
  • PDB/3INT