Localization of Cysteine 302 at the Active Site of Aldehyde Dehydrogenase
The superreactive cysteine was first identified in human cytoplasmic aldehyde dehydrogenase El isozyme, before its primary structure was known, as a part of 35 residue tryptic peptide (Hempel, 1981; Hempel and Pietruszko, 1981; Hempel et al., 1982) by employing iodoacetamide. When the primary structures of the El and E2 isozymes were established (Hempel et al., 1984, 1985; Hsu et al., 1985), this cysteine was found to occupy position 302 in a 500 amino acid residue polypeptide chain. Iodoacetamide fulfilled all criteria for an aldehyde-competitive, active-site-directed reagent with the exception of total inactivation of the mitochondrial E2 isozyme. Since that time, other investigators have also attempted to identify active site residues. Coenzyme-based affinity reagents (von Bahr-Lindstrom et al., 1985) identified cysteines 369 and 302, Nethylmaleimide identified cysteine 49 and 162 (Tu and Weiner, 1988 a,b) and dimethylaminocinnamaldehyde identified serine 74 (Loomes et al., 1990). Our laboratory developed a substrate-based affinity reagent, bromo-acetophenone (MacKerell et al., 1986), which identified glutamate 268 (Abriola et al., 1987).
KeywordsTryptic Peptide Aldehyde Dehydrogenase Aspergillus Nidulans Peptide Mapping Radioactive Label
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- Abriola, D.P., MacKerell, A.D., Jr. and Pietruszko, R., 1990, Correlation of loss of activity of human aldehyde dehydrogenase with reaction of bromoacetophenone with glutamic acid-268 and cysteine-302 residues. Partial-sites reactivity of aldehyde dehydrogenase, Biochem. J., 266:179.PubMedGoogle Scholar
- Blatter, E.E., Tasayco, M.L., Prestwich, G. and Pietruszko, R., 1990, Chemical modification of aldehyde dehydrogenase by a vinyl ketone analog of an insect pheromone, Biochem. J., in press.Google Scholar
- Carey, F.A. and Sundberg, R.J., 1984, Advanced Organic Chemistry. Second Edition. Part A: Structure and Mechanisms, Plenum Press, New York and London, p. 207.Google Scholar
- Fukaya, M., Tayama, K., Tamaki, T., Tagami, H., Okumura, H., Kawamura, Y.. and Beppu, T., 1989, Cloning of the membrane-bound aldehyde dehydrogenase gene of Acetobacter polyoxogenesand improvement of acetic acid production by use of the cloned gene, Appl, Environ. Microbiol., 55:171.Google Scholar
- Hempel, J.D., 1981, Chemical modification of human liver aldehyde dehydrogenase isoenzymes El and E2. Doctoral Dissertation, Rutgers University. Dissertation Abstracts International 42:3664B, University Microfilms No. DA80204216.Google Scholar
- Johansson, J., von Bahr-Lindstrom, Jeck, R., Woenckhaus, C. and Jornvall, H., 1988, Mitochondrial aldehyde dehydrogenase from horse liver. Correlations of the same species variants for both the cytosolic and the mitochondrial forms of an enzyme, Eur. J. Biochem., 172:527.PubMedCrossRefGoogle Scholar
- Prestwich, G.D., Graham, S. McG., Handley, M., Latli, B., Streinz, L. and Tasayco J., M.L., 1988, Enzymatic processing of pheromones and pheromone analogs, Experientia, 45:267.Google Scholar
- Shaw, E., 1970, Chemical modification by active-site-directed reagents. In: The Enzymes (Student Edition), Vol. 1, Structure and Control, Academic Press, p. 91.Google Scholar
- Tasayco J., M.L. and Prestwich, G.D., 1990a, A specific affinity reagent to distinguish aldehyde dehydrogenases and oxidases, J. Biochem. Chem., 265:3094.Google Scholar