title: Isomer-specific retinoic acid biosynthesis in HeLa cells expressing recombinant class I aldehyde dehydrogenases.
abstract: Retinal dehydrogenase type 1 (RALDH1) catalyzes the oxidation of all-trans and 9-cis retinal to the respective retinoic acids (RAs), whereas another member of the aldehyde dehydrogenase family, the phenobarbital-induced aldehyde dehydrogenase (PB-ALDH), is very poorly active. We have previously generated chimeras between these two enzymes that displayed selectivity for retinal isomers in crude bacterial extracts. To examine whether the selectivity of the recombinant enzymes is retained in intact cells, we first assessed whether retinoid-isomerizing activity is present in cultured eukaryotic cells. Our results demonstrate that the only RA isomers detected in RALDH1-expressing or non-expressing cells corresponded to the same steric conformation as the supplied retinoids, indicating a lack of measurable 9-cis/all-trans retinoid-isomerizing activity. Finally, HeLa cells transfected with RALDH1 derivatives that were retinal isomer-selective in vitro produced only the corresponding RA isomers, establishing these enzymes as useful tools to assess the respective roles of the two RA isomers in vivo.
authors.affiliation: Laboratory of Nutrition and Cancer, Centre Hospitalier de l'Université de Montréal-Hotel Dieu, Canada.
authors.forename: Hélène
authors.initials: H
authors.lastname: Brodeur
authors.forename: Maxime
authors.initials: M
authors.lastname: Parisotto
authors.forename: Samuel
authors.initials: S
authors.lastname: Chagnon
authors.forename: Sylvie
authors.initials: S
authors.lastname: Mader
authors.forename: Pangala V
authors.initials: PV
authors.lastname: Bhat
mesh_terms: D000818:Animals; D002460:Cell Line; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D000882:Haplorhini; D006367:HeLa Cells; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics*; D050697:Retinal Dehydrogenase / Q000096:biosynthesis* / Q000145:classification / Q000235:genetics*; D013237:Stereoisomerism; D014162:Transfection; D014212:Tretinoin / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D011993:Recombinant Fusion Proteins; D014212:Tretinoin; D050697:Retinal Dehydrogenase