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PMID:20699116

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Citation

Marchitti, SA, Brocker, C, Orlicky, DJ and Vasiliou, V (2010) Molecular characterization, expression analysis, and role of ALDH3B1 in the cellular protection against oxidative stress. Free Radic. Biol. Med. 49:1432-43

Abstract

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.

Links

PubMed PMC3457645 Online version:10.1016/j.freeradbiomed.2010.08.004

Keywords

Aldehyde Oxidoreductases/genetics; Aldehyde Oxidoreductases/metabolism; Aldehydes/metabolism; Animals; Cytoprotection; Cytosol/metabolism; Epithelial Cells/drug effects; Epithelial Cells/metabolism; Epithelial Cells/pathology; HEK293 Cells; Humans; Lipid Peroxidation/drug effects; Lipid Peroxidation/genetics; Liver/drug effects; Liver/metabolism; Liver/pathology; Mice; Mice, Inbred C57BL; Oxidants/pharmacology; Oxidative Stress; Transgenes/genetics

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

HUMAN:AL3B1

acts_upstream_of_or_within

GO:0046185: aldehyde catabolic process

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:AL3B1

enables

GO:0004030: aldehyde dehydrogenase [NAD(P)+] activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

HUMAN:AL3B1

located_in

GO:0005737: cytoplasm

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:AL3B1

acts_upstream_of_or_within

GO:0034599: cellular response to oxidative stress

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:AL3B1

acts_upstream_of_or_within

GO:0006979: response to oxidative stress

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

MOUSE:AL3B1

located_in

GO:0005829: cytosol

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:AL3B1

enables

GO:0004030: aldehyde dehydrogenase [NAD(P)+] activity

ECO:0000266: sequence orthology evidence used in manual assertion

UniProtKB:P43353

F

Seeded From UniProt

complete


See also

References

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