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. 1967 Jul;104(1):165–172. doi: 10.1042/bj1040165

The specificities and configurations of ternary complexes of yeast and liver alcohol dehydrogenases

F M Dickinson 1, K Dalziel 1
PMCID: PMC1270557  PMID: 4291993

Abstract

1. Some aspects of the substrate specificities of liver and yeast alcohol dehydrogenases have been investigated with pentan-3-ol, heptan-4-ol, (+)-butan-2-ol, (±)-butan-2-ol, (±)-hexan-3-ol and (±)-octan-2-ol as potential substrates. The liver enzyme is active with all substrates tested, including both isomers of each optically active alcohol. In contrast, the yeast enzyme is completely inactive towards those secondary alcohols where both alkyl groups are larger than methyl and active with only the (+)-isomers of butan-2-ol and octan-2-ol. 2. The absence of stereospecificity of liver alcohol dehydrogenase towards optically active secondary alcohols and its broad specificity towards secondary alcohols in general are explained in terms of an alkyl-binding site that will react with a variety of alkyl groups and the ability of the enzyme to accommodate a fairly large unbound alkyl group in an active enzyme–NAD+–secondary alcohol ternary complex. The absolute optical specificity of the yeast enzyme towards n-alkylmethyl carbinols and its unreactivity towards pentan-3-ol, hexan-3-ol and heptan-4-ol are explained by its inability to accept alkyl groups larger than methyl in the unbound position in a viable ternary complex. 3. Comparison of the known configurations of the n-alkylmethyl carbinols and [1-2H]ethanol and [1-3H]geraniol, which have been used in stereospecificity studies with these enzymes by other workers, provides strong evidence for which alkyl group of the substrate is bound to the enzyme in the oxidation of n-alkylmethyl carbinols. The conclusions reached are, for butan-2-ol oxidation with the liver enzyme, confirmed by deductions from kinetic data obtained with (+)-butan-2-ol and a sample of butan-2-ol containing 66% of (−)-butan-2-ol. 4. Initial-rate parameters for the oxidations of (+)-butan-2-ol, 66% (−)-butan-2-ol and pentan-3-ol by NAD with liver alcohol dehydrogenase are presented. The data are completely consistent with a general mechanism of catalysis previously proposed for this enzyme.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BURTON K., WILSON T. H. The free-energy changes for the reduction of diphosphopyridine nucleotide and the dehydrogenation of L-malate and L-glycerol 1-phosphate. Biochem J. 1953 Apr;54(1):86–94. doi: 10.1042/bj0540086. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. CORNFORTH J. W., RYBACK G. Stereochemistry of enzymic hydrogen transfer to pyridine nucleotides. Biochem Biophys Res Commun. 1962 Nov 27;9:371–375. doi: 10.1016/0006-291x(62)90018-9. [DOI] [PubMed] [Google Scholar]
  3. DALZIEL K. Kinetic studies of liver alcohol dehydrogenase. Biochem J. 1962 Aug;84:244–254. doi: 10.1042/bj0840244. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. DALZIEL K. The preparation and properties of crystalline alcohol dehydrogenase from liver. Biochem J. 1961 Aug;80:440–445. doi: 10.1042/bj0800440. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. DALZIEL K. The purification of nicotinamide adenine dinucleotide and kinetic effects of nucleotide impurities. J Biol Chem. 1963 Apr;238:1538–1543. [PubMed] [Google Scholar]
  6. DUGGAN P. F., DONNELLY D. M., MELODY D. P. THE REACTION OF GLYOXYLATE WITH TRIS BUFFER UNDER PHYSIOLOGICAL CONDITIONS. Ir J Med Sci. 1964 Apr;460:163–168. doi: 10.1007/BF02969129. [DOI] [PubMed] [Google Scholar]
  7. Dalziel K., Dickinson F. M. Substrate activation and inhibition in coenzyme-substrate reactions cyclohexanol oxidation catalysed by liver alcohol dehydrogenase. Biochem J. 1966 Aug;100(2):491–500. doi: 10.1042/bj1000491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dalziel K., Dickinson F. M. The kinetics and mechanism of liver alcohol dehydrogenase with primary and secondary alcohols as substrates. Biochem J. 1966 Jul;100(1):34–46. doi: 10.1042/bj1000034. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dickinson F. M., Dalziel K. Substrate specificity and stereospecificity of alcohol dehydrogenases. Nature. 1967 Apr 1;214(5083):31–33. doi: 10.1038/214031a0. [DOI] [PubMed] [Google Scholar]
  10. HARRIS I. STRUCTURE AND CATALYTIC ACTIVITY OF ALCOHOL DEHYDROGENASES. Nature. 1964 Jul 4;203:30–34. doi: 10.1038/203030a0. [DOI] [PubMed] [Google Scholar]
  11. LEVY H. R., VENNESLAND B. The stereospecificity of enzymatic hydrogen transfer from diphosphopyridine nucleotide. J Biol Chem. 1957 Sep;228(1):85–96. [PubMed] [Google Scholar]
  12. MAHLER H. R. The use of amine buffers in studies with enzymes. Ann N Y Acad Sci. 1961 Jun 17;92:426–439. doi: 10.1111/j.1749-6632.1961.tb44992.x. [DOI] [PubMed] [Google Scholar]

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