Human brain "high Km" aldehyde dehydrogenase: purification, characterization, and identification as NAD+ -dependent succinic semialdehyde dehydrogenase.
NAD-dependent succinic semialdehyde dehydrogenase (EC 1.2.1.24) has been purified to homogeneity from human brain via ion-exchange chromatography and affinity chromatography employing Blue Sepharose and 5'-AMP Sepharose. Succinic semialdehyde dehydrogenase was never previously purified to homogeneity from any species; this preparation therefore allows the determination of its molecular weight, subunit molecular weight, subunit composition, isoelectric points, and substrate specificity for the first time. The enzyme is a tetramer of Mr230,000 to 245,000 and consists of weight-nonidentical subunits (Mr 61,000 and 63,000). On isoelectric focusing the enzyme separates into five bands with the following isoelectric points: 6.3, 6.6, 6.8, 6.95, and 7.15. Its substrates include glutaric semialdehyde, nitrobenzaldehyde, and short chain aliphatic aldehydes in addition to succinic semialdehyde which is the best substrate. The Km values for succinic semialdehyde, acetaldehyde, and propionaldehyde are 1,875, and 580 microM, respectively. The enzyme is inactive with 3,4-dihydroxyphenylacetaldehyde and indole-3-acetaldehyde as substrates. Its subcellular localization is in the mitochondrial fraction. Succinic semialdehyde dehydrogenase is sensitive to inhibition by disulfiram (a drug used therapeutically to produce alcohol aversion) resembling, in this respect, aldehyde dehydrogenase (EC 1.2.1.3). It does not, however, interact with the antibody developed in the rabbit vs aldehyde dehydrogenase, suggesting that the two enzymes are structurally distinct.
Ryzlak
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Center of Alcohol Studies, Rutgers University, Piscataway, New Jersey 08855-0969.
Pietruszko
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D000444:Aldehyde Dehydrogenase / Q000037:antagonists & inhibitors / Q000145:classification / Q000302:isolation & purification*; D000445:Aldehyde Oxidoreductases / Q000037:antagonists & inhibitors / Q000145:classification / Q000302:isolation & purification*; D001921:Brain / Q000201:enzymology*; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D004221:Disulfiram / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D007700:Kinetics; D008970:Molecular Weight; D013347:Subcellular Fractions / Q000201:enzymology; D013379:Substrate Specificity; D050604:Succinate-Semialdehyde Dehydrogenase (NADP+)
D000445:Aldehyde Oxidoreductases; D050604:Succinate-Semialdehyde Dehydrogenase (NADP+); D000444:Aldehyde Dehydrogenase; D004221:Disulfiram