title: Enzymatic characteristics of retinal dehydrogenase type I expressed in Escherichia coli.
abstract: We expressed RalDH(I) in Escherichia coli and have shown that it functions in vitro with the complex CRBP-retinal (cellular retinol-binding protein) as substrate, either generated in situ from the complex CRBP-retinol and microsomal retinol dehydrogenases or provided directly as CRBP-retinal. Recombinant RalDH(I) had kinetic constants with CRBP-retinal of: Hill coefficient 1.8; K0.5 0.8 microM; and Vm 1.5 nmol/min/mg of protein at 25 degrees C. Apo-CRBP inhibited the reaction with CRBP-retinal with an IC50 of 1.4 microM. Citral inhibited RalDH(I) with an IC50 of approximately 1 microM compared to an IC50 of approximately 12 microM for RalDH(II), but did not serve as substrate for RalDH(I). RalDH(I) did not catalyze efficiently the dehydrogenation of acetaldehyde, but showed higher Vmax/Km values for hexanal, octanal, decanal and benzaldehyde than for either propanal or retinal. These data extend the characterization of RalDH(I), show that apo-CRBP competes with holo-CRBP as substrate for RalDH(I), and expand insight into the pathways of retinoic acid biogenesis from the most abundant substrates in vivo, retinoid-liganded CRBP.
authors.affiliation: Department of Biochemistry, School of Medicine and Biomedical Sciences, State University of New York at Buffalo 14214, USA.
authors.forename: P
authors.initials: P
authors.lastname: Penzes
authors.forename: X
authors.initials: X
authors.lastname: Wang
authors.forename: J L
authors.initials: JL
authors.lastname: Napoli
mesh_terms: D000445:Aldehyde Oxidoreductases / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D000447:Aldehydes / Q000378:metabolism; D000818:Animals; D003600:Cytosol / Q000201:enzymology; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D008099:Liver / Q000201:enzymology; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry*; D050697:Retinal Dehydrogenase; D013329:Structure-Activity Relationship
chemical_list: D000447:Aldehydes; D011994:Recombinant Proteins; D000445:Aldehyde Oxidoreductases; D050697:Retinal Dehydrogenase