Cloning of a rat cDNA encoding retinal dehydrogenase isozyme type I and its expression in E. coli.
Peptides sequenced from the purified rat liver cytosolic retinal dehydrogenase P1 [Posch, K.C., Burns, R.D. and Napoli, J.L., 1992. Biosynthesis of all-trans-retinoic acid from retinal: recognition of retinal bound to cellular retinol-binding protein (type I) as substrate by a purified cytosolic dehydrogenase. J. Biol. Chem. 267, 19676-19682] were used to design oligonucleotides for cloning its cDNA. The deduced amino acid sequence of P1, now designated retinal dehydrogenase type I or RalDH(I), has close similarity with mouse AHD-2 and rat kidney aldehyde dehydrogenase, but is distinct from rat phenobarbital-inducible aldehyde dehydrogenase (PIADH), the presumed rat liver homolog of mouse AHD-2. Rat kidney (100%) and lung (88%) show relatively high mRNA levels of RalDH(I), liver (34%) and brain (22%) have moderate levels, and testis (8%) has low levels. Retinoid status affects RalDH(I) mRNA levels differently in different tissues. E. coli-expressed RalDH(I) exhibits allosteric kinetics for retinal with a Hill coefficient of 1.7, a K0.5 value of 1.4 microM and a Vmax of 52 nmol min(-1) mg(-1) protein. These data establish the cospecificity of P1 and RalDH(I), show that retinoid status affects expression of its mRNA in a tissue-dependent manner, and illustrate that aldehyde dehydrogenase isozymes with extensive homology can participate in different metabolic paths, e.g., RalDH vs. PIADH.
Penzes
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P
Department of Biochemistry, School of Medicine and Biomedical Sciences, State University of New York at Buffalo, 14214, USA.
Wang
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X
Sperkova
Z
Z
Napoli
J L
JL
D000444:Aldehyde Dehydrogenase / Q000737:chemistry / Q000378:metabolism; D000445:Aldehyde Oxidoreductases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D000494:Allosteric Regulation; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics*; D007668:Kidney / Q000201:enzymology; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D050697:Retinal Dehydrogenase; D013379:Substrate Specificity; D014212:Tretinoin / Q000378:metabolism
D018076:DNA, Complementary; D007527:Isoenzymes; D012333:RNA, Messenger; D011994:Recombinant Proteins; D014212:Tretinoin; D000445:Aldehyde Oxidoreductases; D000444:Aldehyde Dehydrogenase; D050697:Retinal Dehydrogenase