Regulation of alcohol dehydrogenases

The alcohol dehydrogenase (ADH: EC.1.1.1.1) from the fruitfly Drosophila belongs to the Short-chain Dehydrogenase Reductase (SDR) family. This family consists of enzymes from species ranging from bacteria to humans, and they vary largely with respect to the substrates they act on. Among the substrates metabolized by the different SDR enzymes are steroids, prostaglandins, sugars and small alcohols such as ethanol. Common for all these enzymes are that they are not metalloenzymes, and they perform hydrogenation / dehydrogenation reactions of carbonyl / hydroxyl groups in the different substrates, using either NAD(P)H or NAD(P)+ as coenzyme (eqn.1).

Alcohol + NAD(P)+ _ NAD(P)H + H+ + Aldehyde / Ketone (eqn.1)

The active site vary largely between the different SDR enzymes, but they share the same catalytic residues in the active site, and hence, the catalytic mechanism may be identical. X-ray crystallography showed that the enzymes from the SDR family are single domain enzymes. In humans, SDR enzymes are involved in prostaglandin and steroid metabolism, and an altered activity is associated with disease states such as high blood pressure and various types of cancer.

Ongoing and future projects:

(1) Biochemical characterization of Drosophila ADH. Here we focus on the reaction mechanism of the enzyme, chemical function of essential amino acids and the role of a water channel in the molecule, enzyme-substrate and enzyme-inhibitor interactions.

(2) Biochemical characterization of human steroid dehydrogenases. This includes molecular modelling studies and inhibitor design.

(3) Studies of steroid dehydrogenases in isolated cancer cells, tumor tissue and corresponding normal cells and organ cultures. Our intention is to determine if these enzyme activities are important for tumor growth and metastasis, and if so, are they involved in the regulation of matrix metalloproteinases (MMPs) and their inhibitors (TIMPs).


Selected publications:

Benach J, Winberg JO, Svendsen JS, Atrian S, Gonzalez-Duarte R & Ladenstein R.
Drosophila alcohol dehydrogenase: Acetate-enzyme interactions and novel insights into the effects of electrostatics on catalysis.
J Mol Biol 345: 579 - 598, 2005. Abstract

Winberg JO, Brendskag MK, Sylte I, Lindstad RI & McKinley-McKee JS.
The catalytic triad in Drosophila alcohol dehydrogenase: pH, temperature and molecular modelling studies.
J. Mol. Biol. 294: 601-616, 1999. Abstract

Brendskag MK, McKinley-McKee JS & Winberg JO.
Drosophila lebanonensis alcohol dehydrogenase: pH dependence of the kinetic coefficients.
Biochim. Biophys. Acta, 1431: 74-86, 1999. Abstract


Winberg JO & McKinley-McKee JS.
Drosophila melanogaster alcohol dehydrogenase: reaction mechanism of aldehyde oxidation and dismutation.
Biochem J 329: 561-570, 1998. Abstract

Aasmoe L, Winberg JO & Aarbakke J.
The role of alcohol dehydrogenase isoenzymes in the oxidation of glycolethers in male and female rats.
Toxicol. Applied Pharmacol. 150: 86-90, 1998. Abstract

Kvernmo T, Winberg JO & McKinley-McKee JS.
Reversible and irreversible inhibition of sheep liver sorbitol dehydrogenase with Cibacrone Blue 3GA and Eriochrome Black T.
Int J Biochem Cell Biol 28(3): 303-309, 1996. Abstract

Winberg JO & McKinley-McKee JS.
Drosophila melanogaster alcohol dehydrogenase: Product inhibition studies.
Biochem J 301: 901-909, 1994. Abstract

Winberg JO, Martinoni B, Roten C & McKinley-McKee JS.
Drosophila alcohol dehydrogenase: Stereoselective hydrogentransfer from ethanol.
Biochem Mol Biol Int 31: 651-658, 1993. Abstract

Winberg JO & McKinley-McKee JS.
Kinetic Interpretations of active site topologies and residue exchanges in Drosophila alcohol dehydrogenases.
Int J Biochem 24: 169-181, 1992 (Minireview). Abstract