The (2R)-hydroxycarboxylate-viologen-oxidoreductase from Proteus vulgaris is a molybdenum-containing iron-sulphur protein

Eur J Biochem. 1994 Jun 15;222(3):1025-32. doi: 10.1111/j.1432-1033.1994.tb18954.x.

Abstract

An oxidoreductase with an extremely broad substrate specificity reducing reversibly 2-oxocarboxylates at the expense of reduced artificial redox mediators to (2R)-hydroxycarboxylates has been purified to a specific activity of up to 1800 mumol.min-1.mg-1 for the reduction of phenylpyruvate. The membrane-bound non-pyridine nucleotide-dependent enzyme appears in the form of various oligomers of the 80-kDa monomer. The isoelectric point is 5.1. Based on a molecular mass of 80 kDa the enzyme contains up to one molybdenum, four iron and four sulphur atoms. After growth on 99Mo-labelled molybdate, enzyme and radioactivity coincided as shown by gel electrophoresis. Permanganate oxidation delivers 0.7 mol pterin-6-carboxylic acid. The molybdenum cofactor is a mononucleotide. The enzyme is inhibited by cyanide. The first 20 amino acids have been determined. The enzyme belongs to the rare group of molybdoenzymes which possess no further prosthetic groups than the iron-sulphur clusters.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Amino Acids / analysis
  • Bacterial Proteins*
  • Coenzymes*
  • Electrophoresis, Polyacrylamide Gel
  • Iron-Sulfur Proteins / chemistry
  • Iron-Sulfur Proteins / isolation & purification*
  • Iron-Sulfur Proteins / metabolism
  • Isoelectric Point
  • Metalloproteins / analysis
  • Metalloproteins / chemistry
  • Molecular Sequence Data
  • Molecular Weight
  • Molybdenum / analysis*
  • Molybdenum / metabolism
  • Molybdenum Cofactors
  • Oxidation-Reduction
  • Oxidoreductases / chemistry
  • Oxidoreductases / isolation & purification*
  • Oxidoreductases / metabolism
  • Proteus vulgaris / enzymology*
  • Pteridines / analysis
  • Pteridines / chemistry
  • Pterins / analysis
  • Substrate Specificity
  • Viologens / metabolism

Substances

  • Amino Acids
  • Bacterial Proteins
  • Coenzymes
  • Iron-Sulfur Proteins
  • Metalloproteins
  • Molybdenum Cofactors
  • Pteridines
  • Pterins
  • Viologens
  • Molybdenum
  • molybdenum cofactor
  • Oxidoreductases
  • HVOR protein, Proteus vulgaris