Succinic semialdehyde dehydrogenase from mammalian brain: subunit analysis using polyclonal antiserum.
1. NAD(+)-dependent succinic semialdehyde dehydrogenase was purified to apparent homogeneity from rat brain and highly purified from human brain. 2. Molecular exclusion chromatography of the purified enzymes on Sephadex G-150 and G-200 revealed M(r) values of 203,000 and 191,000 for rat and human, respectively. 3. Electrophoresis on sodium dodecylsulfate polyacrylamide gels revealed a single subunit of M(r) 54,000 for rat and 58,000 for human. Isoelectric focusing of the purified rat enzyme yielded a pI of 6.1. 4. For both proteins, Km values for short-chain aldehydes acetaldehyde and propionaldehyde ranged from 0.33 to 2.5 mM; Km values for succinic semialdehyde were in the 2-4 microM range. 5. The subunit structure of both enzymes was investigated in brain extracts and purified preparations by immunoblotting, using a polyclonal rabbit antiserum against the purified rat brain enzyme. 6. For rat and human extracts, single bands were detected at M(r) 54,000 and 58,000, comparable to findings in the purified preparations. Immunoblotting analyses in other species (guinea pig, hamster, mouse and rabbit) revealed single subunits of M(r) 54,000-56,500.
Chambliss
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KL
Kimberly H. Courtwright and Joseph W. Summers Metabolic Disease Center, Baylor University Medical Center, Dallas, TX 75226.
Gibson
K M
KM
D000445:Aldehyde Oxidoreductases / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D015153:Blotting, Western; D001921:Brain / Q000201:enzymology*; D004789:Enzyme Activation; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007106:Immune Sera; D007525:Isoelectric Focusing; D007700:Kinetics; D008970:Molecular Weight; D051381:Rats; D013045:Species Specificity; D013379:Substrate Specificity; D050644:Succinate-Semialdehyde Dehydrogenase
D007106:Immune Sera; D000445:Aldehyde Oxidoreductases; C493643:ALDH5A1 protein, human; C493647:Aldh5a1 protein, rat; D050644:Succinate-Semialdehyde Dehydrogenase