Comparative roles of histidine-51 in human β1β1 and threonine-51 in ππ alcohol dehydrogenases

Gerard J. Davis, Lucinda G. Carr, Thomas D. Hurley, Ting Kai Li, William F. Bosron

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20 Scopus citations


Histidine at position 51 of the class I β1 alcohol dehydrogenase (ADH) functions as a general base by indirectly abstracting a proton from the alcohol substrate through a hydrogen-bonded proton relay system. The human class II π-ADH was reported to be polymorphic, having either Ser or Thr, but not His at position 51. It is unknown whether Ser or Thr51 have a catalytic role in ethanol oxidation with π-ADH. Accordingly, we expressed and purified recombinant mutants of π-ADH with Thr, Ser, and His at position 51. At pH 6.5, values for Vmax/Km for ethanol were 0.30, 0.10, and 0.09 min-1 mM-1 for π51Thr, π51Ser, and π51His ADH, respectively. Hence the effects of the substitutions were much less than the 11-fold decrease in Vmax/Km observed for β1-ADH when a neutral amino acid (Gln) was substituted for His51. Addition of a buffer base (400 mM glycylglycine) had little effect on Vmax/Km of recombinant π51Thr or π51Ser ADH, while it increased Vmax/Km for ethanol 7-fold for the β151Gln ADH. We conclude that there is no evidence for Thr51 of π-ADH participating in a proton relay similar to that seen in β1-ADH and that a base at position 51 may not be a universal requirement for a functional alcohol dehydrogenase with a moderate efficiency for ethanol oxidation at a physiological pH.

Original languageEnglish (US)
Pages (from-to)307-312
Number of pages6
JournalArchives of Biochemistry and Biophysics
Issue number2
StatePublished - Jun 1994

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

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