The overall goal of this project is to determine the role of cooperativity in liver alcohol dehydrogenase. Liver alcohol dehydrogenase is a rate-limiting factor in ethanol metabolism, underscoring its importance in ethanol elimination. Horse and human liver alcohol dehydrogenases exist as a mixture of homo-and heterodimeric isoenzymes. Apparent cooperativity has been observed in many of these isoenzymes, specifically for the gamma2gamma2 human enzyme and the heterodimeric isoenzymes from both horse and human liver tissues. The steady-state and transient kinetics of recombinant gamma2gamma2 human enzyme will be determined in an effort to understand the observed cooperativity. Along with these studies, the binding of substrates and coenzymes to gamma2gamma2 human enzyme will be measured using epr and fluorescence spectroscopy. Site-directed mutagenesis will be used to disrupt coenzyme binding to determine if the conformational change associated with coenzyme binding to gamma2gamma2 is involved in mediating the subunit interactions. Cooperativity in the alphagamma2 human liver isoenzyme and in horse liver heterodimers consisting of a wild-type and a mutant subunit will also be studied. The results of this research will contribute to the current understanding of ethanol metabolism in man.

Agency
National Institute of Health (NIH)
Institute
National Institute on Alcohol Abuse and Alcoholism (NIAAA)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
5F32AA005530-02
Application #
2893984
Study Section
Special Emphasis Panel (ZRG4-ALTX-1 (01))
Program Officer
Isaki, Leslie
Project Start
1999-08-17
Project End
Budget Start
1999-08-17
Budget End
2000-08-16
Support Year
2
Fiscal Year
1999
Total Cost
Indirect Cost
Name
University of Iowa
Department
Biochemistry
Type
Schools of Medicine
DUNS #
041294109
City
Iowa City
State
IA
Country
United States
Zip Code
52242