Composition of the Coenzyme F420-dependent Formate Dehydrogenase From Methanobacterium Formicicum

J Bacteriol. 1986 Feb;165(2):405-11. doi: 10.1128/jb.165.2.405-411.1986.

Abstract

The coenzyme F420-dependent formate dehydrogenase from Methanobacterium formicicum was purified to electrophoretic homogeneity by anoxic procedures which included the addition of azide, flavin adenine dinucleotide (FAD), glycerol, and 2-mercaptoethanol to all buffer solutions to stabilize activity. The enzyme contains, in approximate molar ratios, 1 FAD molecule and 1 molybdenum, 2 zinc, 21 to 24 iron, and 25 to 29 inorganic sulfur atoms. Denaturation of the enzyme released a molybdopterin cofactor. The enzyme has a molecular weight of 177,000 and consists of one each of two different subunits, giving the composition alpha 1 beta 1. The molecular weight of the alpha-subunit is 85,000, and that of the beta-subunit is 53,000. The UV-visible spectrum is typical of nonheme iron-sulfur flavoprotein. Reduction of the enzyme facilitated dissociation of FAD, and the FAD-depleted enzyme was unable to reduce coenzyme F420. Preincubation of the FAD-depleted enzyme with FAD restored coenzyme F420-dependent activity.

Publication types

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

MeSH terms

  • Aldehyde Oxidoreductases / isolation & purification*
  • Euryarchaeota / enzymology*
  • Flavin-Adenine Dinucleotide / metabolism
  • Formate Dehydrogenases / isolation & purification*
  • Iron / analysis
  • Macromolecular Substances
  • Metalloproteins / isolation & purification*
  • Molecular Weight
  • Molybdenum / analysis
  • Riboflavin / analogs & derivatives
  • Riboflavin / metabolism
  • Spectrum Analysis
  • Sulfur / analysis
  • Zinc / analysis

Substances

  • Macromolecular Substances
  • Metalloproteins
  • Flavin-Adenine Dinucleotide
  • coenzyme F420
  • Sulfur
  • Molybdenum
  • Iron
  • Formate Dehydrogenases
  • Aldehyde Oxidoreductases
  • Zinc
  • Riboflavin