Aldehyde Dehydrogenase 1B1: Molecular Cloning and Characterization of a Novel Mitochondrial Acetaldehyde-Metabolizing Enzyme
- 1 October 2010
- journal article
- Published by Elsevier in Drug Metabolism and Disposition
- Vol. 38 (10) , 1679-1687
- https://doi.org/10.1124/dmd.110.034678
Abstract
Ethanol-induced damage is largely attributed to its toxic metabolite, acetaldehyde. Clearance of acetaldehyde is achieved by its oxidation, primarily catalyzed by the mitochondrial class II aldehyde dehydrogenase (ALDH2). ALDH1B1 is another mitochondrial aldehyde dehydrogenase (ALDH) that shares 75% peptide sequence homology with ALDH2. Recent population studies in whites suggest a role for ALDH1B1 in ethanol metabolism. However, to date, no formal documentation of the biochemical properties of ALDH1B1 has been forthcoming. In this current study, we cloned and expressed human recombinant ALDH1B1 in Sf9 insect cells. The resultant enzyme was purified by affinity chromatography to homogeneity. The kinetic properties of purified human ALDH1B1 were assessed using a wide range of aldehyde substrates. Human ALDH1B1 had an exclusive preference for NAD+ as the cofactor and was catalytically active toward short- and medium-chain aliphatic aldehydes, aromatic aldehydes, and the products of lipid peroxidation, 4-hydroxynonenal and malondialdehyde. Most importantly, human ALDH1B1 exhibited an apparent Km of 55 μM for acetaldehyde, making it the second low Km ALDH for metabolism of this substrate. The dehydrogenase activity of ALDH1B1 was sensitive to disulfiram inhibition, a feature also shared with ALDH2. The tissue distribution of ALDH1B1 in C57BL/6J mice and humans was examined by quantitative polymerase chain reaction, Western blotting, and immunohistochemical analysis. The highest expression occurred in the liver, followed by the intestinal tract, implying a potential physiological role for ALDH1B1 in these tissues. The current study is the first report on the expression, purification, and biochemical characterization of human ALDH1B1 protein.This publication has 46 references indexed in Scilit:
- Organ-Specific Roles of CYP1A1 during Detoxication of Dietary Benzo[a]pyreneMolecular Pharmacology, 2010
- Corneal aldehyde dehydrogenases: Multiple functions and novel nuclear localizationBrain Research Bulletin, 2010
- Genetic determinants of both ethanol and acetaldehyde metabolism influence alcohol hypersensitivity and drinking behaviour among ScandinaviansClinical and Experimental Allergy, 2009
- Inhibition of hepatic mitochondrial aldehyde dehydrogenase by carbon tetrachloride through JNK-mediated phosphorylationFree Radical Biology & Medicine, 2009
- Human aldehyde dehydrogenase genes: alternatively spliced transcriptional variants and their suggested nomenclaturePharmacogenetics and Genomics, 2009
- The Association of ADH and ALDH Gene Variants With Alcohol Drinking Habits and Cardiovascular Disease Risk FactorsAlcohol, Clinical and Experimental Research, 2008
- Activation of Aldehyde Dehydrogenase-2 Reduces Ischemic Damage to the HeartScience, 2008
- Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2−ΔΔCT MethodMethods, 2001
- Membrane Topology and Retention of Microsomal Aldehyde Dehydrogenase in the Endoplasmic ReticulumJournal of Biological Chemistry, 1996
- The Novel Aldehyde Dehydrogenase Gene, ALDH5, Encodes an Active Aldehyde Dehydrogenase EnzymeBiochemical and Biophysical Research Communications, 1995