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The Biochemical journal1987; 242(2); 499-503; doi: 10.1042/bj2420499

Inactivation of horse liver mitochondrial aldehyde dehydrogenase by disulfiram. Evidence that disulfiram is not an active-site-directed reagent.

Abstract: The inhibition of mitochondrial (pI 5) horse liver aldehyde dehydrogenase by disulfiram (tetraethylthiuram disulphide) was investigated to determine if the drug was an active-site-directed inhibitor. Stoichiometry of inhibition was determined by using an analogue, [35S]tetramethylthiuram disulphide. A 50% loss of the dehydrogenase activity was observed when only one site per tetrameric enzyme was modified, and complete inactivation was not obtained even after seven sites per tetramer were modified. Modification of only two sites accounted for a loss of 75% of the initial catalytic activity. The number of functioning active sites per tetrameric enzyme, as determined by the magnitude of the pre-steady-state burst of NADH formation, did not decrease until approx. 75% of the catalytic activity was lost. These data indicate that disulfiram does not modify the essential nucleophilic amino acid at the active site of the enzyme. The data support an inactivation mechanism involving the formation of a mixed disulphide with a non-essential cysteine residue, resulting in a lowered specific activity of the enzyme.
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Publication Date: 1987-03-01 PubMed ID: 3593264PubMed Central: PMC1147733DOI: 10.1042/bj2420499Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research investigates how the drug disulfiram inhibits mitochondrial horse liver aldehyde dehydrogenase, concluding that the drug does not target the active site of the enzyme directly. Rather, it interacts with a non-essential cysteine residue, leading to decreased enzyme activity.

Background of the Study

  • The study revolves around the interaction between a drug called disulfiram and an enzyme present in horse liver known as mitochondrial aldehyde dehydrogenase.
  • Disulfiram’s role as an inhibitive agent for mitochondrial aldehyde dehydrogenase was under scrutiny with a particular interest in understanding whether it inhibits the enzyme by directly targeting its active site.

Method and Investigation

  • The researchers used [35S]tetramethylthiuram disulphide, which is an analogue of disulfiram, to help determine the stoichiometry (the measurement of quantitative relationship) of inhibition.
  • By examining the enzyme’s reaction to disulfiram, they found out the extent of loss in dehydrogenase activity with each modification of the enzyme’s sites.

Key Findings

  • The team observed that with the modification of only one site per tetrameric enzyme, there was a 50% loss in dehydrogenase activity.
  • Interestingly, complete inactivation did not occur even after modification of seven sites per tetramer. However, a loss of 75% of the initial catalytic activity was noticed when two sites were modified.
  • Moreover, the number of active sites per tetrameric enzyme did not decrease until approximately 75% of the catalytic activity was lost.

Conclusion

  • The results suggest that disulfiram does not directly modify the essential nucleophilic amino acid at the enzyme’s active site.
  • Rather, the evidence indicates an inactivation mechanism where the drug forms a mixed disulphide with a non-essential cysteine residue. This interaction results in the specific activity of the enzyme being lowered, rather than the enzyme’s complete inactivation.

Cite This Article

APA
Sanny CG, Weiner H. (1987). Inactivation of horse liver mitochondrial aldehyde dehydrogenase by disulfiram. Evidence that disulfiram is not an active-site-directed reagent. Biochem J, 242(2), 499-503. https://doi.org/10.1042/bj2420499

Publication

The Biochemical journal
ISSN: 0264-6021
NlmUniqueID: 2984726R
Country: England
Language: English
Volume: 242
Issue: 2
Pages: 499-503

Researcher Affiliations

Sanny, C G
    Weiner, H

      MeSH Terms

      • Aldehyde Dehydrogenase / antagonists & inhibitors
      • Animals
      • Binding Sites / drug effects
      • Disulfiram / pharmacology
      • Horses
      • Kinetics
      • Magnesium / pharmacology
      • Mitochondria, Liver / enzymology
      • Thiram / pharmacology

      Grant Funding

      • AA05812 / NIAAA NIH HHS
      • K05-AA00028 / NIAAA NIH HHS

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      Citations

      This article has been cited 3 times.
      1. Marikovsky M. Thiram inhibits angiogenesis and slows the development of experimental tumours in mice. Br J Cancer 2002 Mar 4;86(5):779-87.
        doi: 10.1038/sj.bjc.6600078pubmed: 11875743google scholar: lookup
      2. Lee BH, Song YS, Park J, Ryu JC. Metabolism and pharmacokinetics of S-(N,N-diethyldithiocarbamoyl)-N-acetyl-L-cysteine in rats. Arch Pharm Res 1994 Dec;17(6):428-33.
        doi: 10.1007/BF02979120pubmed: 10319153google scholar: lookup
      3. Kitson TM. Effect of disulfiram on the pre-steady-state burst in the reactions of sheep liver cytoplasmic aldehyde dehydrogenase. Biochem J 1987 Dec 15;248(3):989-91.
        doi: 10.1042/bj2480989pubmed: 3435497google scholar: lookup
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