title: Aldehyde dehydrogenase 7A1 (ALDH7A1) attenuates reactive aldehyde and oxidative stress induced cytotoxicity.
abstract: Mammalian aldehyde dehydrogenase 7A1 (ALDH7A1) is homologous to plant ALDH7B1 which protects against various forms of stress such as increased salinity, dehydration and treatment with oxidants or pesticides. Deleterious mutations in human ALDH7A1 are responsible for pyridoxine-dependent and folinic acid-responsive seizures. In previous studies, we have shown that human ALDH7A1 protects against hyperosmotic stress presumably through the generation of betaine, an important cellular osmolyte, formed from betaine aldehyde. Hyperosmotic stress is coupled to an increase in oxidative stress and lipid peroxidation (LPO). In this study, cell viability assays revealed that stable expression of mitochondrial ALDH7A1 in Chinese hamster ovary (CHO) cells provides significant protection against treatment with the LPO-derived aldehydes hexanal and 4-hydroxy-2-nonenal (4HNE) implicating a protective function for the enzyme during oxidative stress. A significant increase in cell survival was also observed in CHO cells expressing either mitochondrial or cytosolic ALDH7A1 treated with increasing concentrations of hydrogen peroxide (H(2)O(2)) or 4HNE, providing further evidence for anti-oxidant activity. In vitro enzyme activity assays indicate that human ALDH7A1 is sensitive to oxidation and that efficiency can be at least partially restored by incubating recombinant protein with the thiol reducing agent β-mercaptoethanol (BME). We also show that after reactivation with BME, recombinant ALDH7A1 is capable of metabolizing the reactive aldehyde 4HNE. In conclusion, ALDH7A1 mechanistically appears to provide cells protection through multiple pathways including the removal of toxic LPO-derived aldehydes in addition to osmolyte generation.
authors.affiliation: Department of Pharmaceutical Sciences, University of Colorado Denver, Aurora, CO 80045, USA.
authors.forename: Chad
authors.initials: C
authors.lastname: Brocker
authors.forename: Miriam
authors.initials: M
authors.lastname: Cantore
authors.forename: Paola
authors.initials: P
authors.lastname: Failli
authors.forename: Vasilis
authors.initials: V
authors.lastname: Vasiliou
mesh_terms: D015074:2-Aminoadipic Acid / Q000031:analogs & derivatives / Q000633:toxicity; D000444:Aldehyde Dehydrogenase / Q000235:genetics / Q000378:metabolism*; D000447:Aldehydes / Q000633:toxicity*; D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity; D015227:Lipid Peroxidation / Q000187:drug effects; D008623:Mercaptoethanol / Q000494:pharmacology; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D016877:Oxidants / Q000633:toxicity; D018384:Oxidative Stress / Q000187:drug effects*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D000447:Aldehydes; D016877:Oxidants; D011994:Recombinant Proteins; D015074:2-Aminoadipic Acid; C000061:allysine; D008623:Mercaptoethanol; D006861:Hydrogen Peroxide; C089507:ALDH7A1 protein, human; D000444:Aldehyde Dehydrogenase; C027576:4-hydroxy-2-nonenal