On the Interaction of Human Liver Aldehyde Dehydrogenase E1 Isoenzyme with Disulfiram and Iodoacetamide

  • John D. Hempel
  • Robert C. Vallari
  • Regina Pietruszko

Abstract

The El isoenzyme of human liver aldehyde dehydrogenase (Km acetaldehyde = 30uM, pH 7.0), when incubated with disulfiram at a stoichiometry of four moles disulfiram/tetrameric El, is immediately inhibited to within 10% of control activity. The inhibition is reversed by 0.1% (v/v) mercaptoethanol, indicating disulfide bridge formation. An indirect attempt to locate, on maps, a peptide binding disulfiram has yielded inconsistent results. Iodoacetamide inhibits El slowly; inhibition is facilitated in the presence of NAD, resulting in loss of ca. 90% of control activity. Incubation with 14C iodoacetamide labels a 16,000 dalton CNBr peptide, and a ca. 4,000 dalton tryptic cleavage product. These fragments can be equated with those which have been suggested by disulfiram.

Keywords

Cellulose Aldehyde Pyridine Trypsin Disulfide 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Blair, A. H. and Bodley, F. H., 1969, Human liver aldehyde dehydrogenase: partial purification and properties, Canad. J. Biochem., 47:265.Google Scholar
  2. Bonner, W. and Laskey, R. A., 1974, A film detection method for 3H labelled proteins and nucleic acids in polyacrylamide gels, Eur. J. Biochem., 46:83.Google Scholar
  3. Deitrich, R. A., Hellerman, L. and Wein, J., 1962, Diphosphopyridinenucleotide-linked aldehyde dehydrogenase (1): specificity and sigma-rho function, J. Biol. Chem., 237: 560.Google Scholar
  4. Deitrich, R. A. and Hellerman, L., 1963, Diphosphopyridine-nucleotide linked aldehyde dehydrogenase (11): inhibitors, J. Biol. Chem., 238:1683.Google Scholar
  5. Deitrich, R. A., 1967, Diphosphopyridine nucleotide-linked aldehyde dehydrogenase (111): sulfhydryl characteristics of the enzyme, Arch. Biochem. Biophys., 119:253.Google Scholar
  6. Deitrich, R. A. and Erwin, V. G., 1971, Mechanism of inhibition of aldehyde dehydrogenase in vivo by disulfiram and diethyldithiocarbamate, Mol. Pharmacol., 7:301.Google Scholar
  7. Faiman, M. D., Dodd, D. E., Nolan, R. J. and Hanzlik, R., 1976, Deter- mination of disulfiram and metabolites in plasma and tissues, in: “Alcohol and Aldehyde Metabolizing Systems,” III, R. G. Thurman, J. R. Williamson, H. R. Drott and B. Chance, eds., Academic Press, New York.Google Scholar
  8. Feldman, R. and Weiner, H., 1972a, Horse liver aldehyde dehydrogenase I: purification and characterization, J. Biol. Chem., 247: 260.Google Scholar
  9. Feldman, R. and Weiner, H., 1972b, Horse liver aldehyde dehydrogenase II: kinetics and mechanistic implications of the dehydrogenase and esterase activity, J. Biol. Chem., 247: 267.Google Scholar
  10. Greenfield, N. J. and Pietruszko, R., 1977, Two aldehyde dehydrogenases from human liver. Isolation via affinity chromatography and characterization of the isoenzymes, Biochem Biophys Acta, 483: 35Google Scholar
  11. Hart, G. J. and Dickinson, F. M., 1977, Some properties of aldehyde dehydrogenase from sheep liver mitochondria, Biochem. J., 163: 261.Google Scholar
  12. Hempel, J. D. and Pietruszko, R., 1978, Structural distinctness of isoenzymes of human liver aldehyde dehydrogenase, Fed. Proc., 37:1798 Abs.Google Scholar
  13. Hirs, C. H. W., 1967, Reduction and S-Carboxymethylation of proteins, in: “Methods in Enzymology,” XI, C. H. W. Hirs, ed., Academic Press, New York.Google Scholar
  14. Jackoby, W. B., 1963, Aldehyde dehydrogenases, in: “The Enzymes,” VII, P. D. Boyer, H. Lardy and K. Myrback, eds., Academic Press, New York.Google Scholar
  15. Kitson, T. M., 1975, The effect of disulfiram on the aldehyde dehydrogeneses of sheep liver, Biochem. J., 151: 407.Google Scholar
  16. Kitson, T. M., 1976, The effect of some analogs of disulfiram on the aldehyde dehydrogenases of sheep liver, Biochem. J., 155: 445.Google Scholar
  17. Kitson, T. M., 1977, The disulfiram-ethanol reaction; a review, J. Stud. Alc., 38:96.Google Scholar
  18. Kitson, T. M., 1978, Studies on the interaction between disulfiram and sheep liver cytoplasmic aldehyde dehydrogenase, Biochem. J. 175:83.Google Scholar
  19. Kraemer, R. J. and Deitrich, R. A., 1968, Isolation and characterization of human liver aldehyde dehydrogenase, J. Biol. Chem., 243:6402.Google Scholar
  20. Laemmli, U. K., 1970, Cleavage of structural proteins during the assembly of the head of bacteriophage T4, Nature, 227: 680.Google Scholar
  21. Racker, E., 1955, Actions and properties of pyridine-nucleotide linked enzymes, Physiol. Rev., 35:1.Google Scholar
  22. Stromme, J. H., 1965, Metabolism of disulfiram and diethydithiocarbamate in rats, Biochem. Pharmacol., 14:393.Google Scholar
  23. Weber, K. and Osborn, M., 1969, The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis, J. Biol. Chem., 244:4406.Google Scholar

Copyright information

© Springer Science+Business Media New York 1980

Authors and Affiliations

  • John D. Hempel
    • 1
  • Robert C. Vallari
    • 1
  • Regina Pietruszko
    • 1
  1. 1.Center of Alcohol StudiesRutgers UniversityNew BrunswickUSA

Personalised recommendations