Molecular characterization, expression analysis, and role of ALDH3B1 in the cellular protection against oxidative stress.
Aldehyde dehydrogenase (ALDH) enzymes are critical in the detoxification of aldehydes. The human genome contains 19 ALDH genes, mutations in which are the basis of several diseases. The expression, subcellular localization, enzyme kinetics, and role of ALDH3B1 in aldehyde- and oxidant-induced cytotoxicity were investigated. ALDH3B1 was purified from Sf9 cells using chromatographic methods, and enzyme kinetics were determined spectrophotometrically. ALDH3B1 demonstrated high affinity for hexanal (K(m)=62 μM), octanal (K(m)=8 μM), 4-hydroxy-2-nonenal (4HNE; K(m)=52 μM), and benzaldehyde (K(m)=46 μM). Low affinity was seen toward acetaldehyde (K(m)=23.3 mM), malondialdehyde (K(m)=152 mM), and the ester p-nitrophenyl acetate (K(m)=3.6 mM). ALDH3B1 mRNA was abundant in testis, lung, kidney, and ovary. ALDH3B1 protein was highly expressed in these tissues and the liver. Immunofluorescence microscopy of ALDH3B1-transfected human embryonic kidney (HEK293) cells and subcellular fractionation of mouse kidney and liver revealed a cytosolic protein localization. ALDH3B1-transfected HEK293 cells were significantly protected from the lipid peroxidation-derived aldehydes trans-2-octenal, 4HNE, and hexanal and the oxidants H(2)O(2) and menadione. In addition, ALDH3B1 protein expression was up-regulated by 4HNE in ARPE-19 cells. The results detailed in this study support a pathophysiological role for ALDH3B1 in protecting cells from the damaging effects of oxidative stress.
Marchitti
Satori A
SA
Molecular Toxicology and Environmental Health Sciences Program, Department of Pharmaceutical Sciences, University of Colorado at Denver, Aurora, CO 80045, USA.
Brocker
Chad
C
Orlicky
David J
DJ
Vasiliou
Vasilis
V
D000445:Aldehyde Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D000447:Aldehydes / Q000378:metabolism; D000818:Animals; D019610:Cytoprotection; D003600:Cytosol / Q000378:metabolism; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D057809:HEK293 Cells; D006801:Humans; D015227:Lipid Peroxidation / Q000187:drug effects / Q000235:genetics; D008099:Liver / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D008810:Mice, Inbred C57BL; D016877:Oxidants / Q000494:pharmacology; D018384:Oxidative Stress; D019076:Transgenes / Q000235:genetics
D000447:Aldehydes; D016877:Oxidants; D000445:Aldehyde Oxidoreductases; C520012:ALDH3B1 protein, human