STRUCTURE OF ALCOHOL AND ALDEHYDE DEHYDROGENASES

Project: Research project

Description

The long-term objective of this research project is to determine the three-
dimensional structure of the multiple molecular forms of the two principal
enzymes involved in the metabolism of ethanol; alcohol dehydrogenase and
aldehyde dehydrogenase. The hypothesis is that knowledge of the three-
dimensional structure of the substrate binding sites of the various
isoenzymes will allow the prediction of their substrate specificities as
well as their contributions to the metabolism of ethanol. Specifically, I
will crystallize and determine the three-dimensional structures of the
human beta1beta1, alphaalpha, pipi, and chichi alcohol dehydrogenase
enzymes complexed with alcohol substrates and the mitochondrial form of
aldehyde dehydrogenase complexed with the coenzyme NAD(H). The recombinant
proteins, expressed and purified from E. coli, will be used for this
purpose to alleviate problems associated with minor contamination of
closely related isoenzyme forms. These forms of human alcohol
dehydrogenase possess amino acid substitutions within their substrate
binding sites and it is the objective of this proposal to evaluate the
effects of these substitutions on substrate specificity and maximal
catalytic activity. A three-dimensional structure for the mitochondrial
form of aldehyde dehydrogenase or closely related enzymes is not available.
Accordingly, little is known about the determinants of catalytic activity
or substrate specificity for the aldehyde dehydrogenase isoenzymes. The
elucidation of its three-dimensional structure by X-ray crystallography
will provide the necessary structural model for determining these
functional relationships.
StatusFinished
Effective start/end date7/1/926/30/97

Funding

  • National Institutes of Health
  • National Institutes of Health: $75,905.00
  • National Institutes of Health: $67,557.00
  • National Institutes of Health: $74,871.00
  • National Institutes of Health

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Aldehyde Dehydrogenase
Alcohol Dehydrogenase
Substrate Specificity
Isoenzymes
Ethanol
Molecular Conformation
Alcohols
Oxidoreductases
Structural Models
Coenzymes
Amino Acid Substitution
NAD
Binding Sites
Enzymes
Escherichia coli
X-Rays
Research
X Ray Crystallography
Recombinant Proteins

ASJC

  • Medicine(all)