Purification and characterization of NAD and NADP-linked alpha-ketoaldehyde dehydrogenases involved in catalyzing the oxidation of methylglyoxal to pyruvate.
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.
D000445:Aldehyde Oxidoreductases / Q000302:isolation & purification* / Q000378:metabolism; D000818:Animals; D006041:Goats; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D046911:Macromolecular Substances; D008970:Molecular Weight; D013439:Sulfhydryl Reagents / Q000494:pharmacology
D046911:Macromolecular Substances; D013439:Sulfhydryl Reagents; D000445:Aldehyde Oxidoreductases; C021928:2-oxoaldehyde dehydrogenase (NAD+)