Purification and Characterization of NAD and NADP-linked Alpha-Ketoaldehyde Dehydrogenases Involved in Catalyzing the Oxidation of Methylglyoxal to Pyruvate

J Biol Chem. 1982 Sep 25;257(18):10566-70.

Abstract

Two alpha-ketoaldehyde dehydrogenases, one catalyzing the oxidation of methylglyoxal to pyruvate with NAD and the other with NADP, were isolated from goat liver and happened to be co-purified. Both the enzymes had been extensively purified to the point where only these two enzymes were present. By affinity chromatography on a thiol-Sepharose column, the two enzymes were separated. Molecular weight of both the enzymes was found to be 42,000 by gel filtration in a Sephadex G-200 column. Electrophoresis on sodium dodecyl sulfate-polyacrylamide gel revealed that the enzymes are composed of single subunits. Interaction with mercurials indicated the presence of SH group(s) in the active site of the NAD-linked enzyme only.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aldehyde Oxidoreductases / isolation & purification*
  • Aldehyde Oxidoreductases / metabolism
  • Animals
  • Goats
  • Kinetics
  • Liver / enzymology*
  • Macromolecular Substances
  • Molecular Weight
  • Sulfhydryl Reagents / pharmacology

Substances

  • Macromolecular Substances
  • Sulfhydryl Reagents
  • Aldehyde Oxidoreductases
  • 2-oxoaldehyde dehydrogenase (NAD+)