Identification and characterization of a stereospecific human enzyme that catalyzes 9-cis-retinol oxidation. A possible role in 9-cis-retinoic acid formation.
All-trans- and 9-cis-retinoic acid are active retinoids for regulating expression of retinoid responsive genes, serving as ligands for two classes of ligand-dependent transcription factors, the retinoic acid receptors and retinoid X receptors. Little is known, however, regarding 9-cis-retinoic acid formation. We have obtained a 1.4-kilobase cDNA clone from a normalized human breast tissue library, which when expressed in CHO cells encodes a protein that avidly catalyzes oxidation of 9-cis-retinol to 9-cis-retinaldehyde. This protein also catalyzes oxidation of 13-cis-retinol at a rate approximately 10% of that of the 9-cis isomer but does not catalyze all-trans-retinol oxidation. NAD+ was the preferred electron acceptor for oxidation of 9-cis-retinol, although NADP+ supported low rates of 9-cis-retinol oxidation. The rate of 9-cis-retinol oxidation was optimal at pHs between 7.5 and 8. Sequence analysis indicates that the cDNA encodes a protein of 319 amino acids that resembles members of the short chain alcohol dehydrogenase protein family. mRNA for the protein is most abundant in human mammary tissue followed by kidney and testis, with lower levels of expression in liver, adrenals, lung, pancreas, and skeletal muscle. We propose that this cDNA encodes a previously unknown stereospecific enzyme, 9-cis-retinol dehydrogenase, which probably plays a role in 9-cis-retinoic acid formation.
Mertz
J R
JR
Department of Cell Biology and Anatomical Sciences, City University of New York Medical School, New York, New York 10031, USA.
Shang
E
E
Piantedosi
R
R
Wei
S
S
Wolgemuth
D J
DJ
Blaner
W S
WS
D000429:Alcohol Oxidoreductases / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D000077556:Alitretinoin; D000595:Amino Acid Sequence; D001483:Base Sequence; D001940:Breast / Q000201:enzymology*; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004822:Epididymis / Q000201:enzymology; D005260:Female; D015723:Gene Library; D006801:Humans; D007668:Kidney / Q000201:enzymology; D008099:Liver / Q000201:enzymology; D008297:Male; D008969:Molecular Sequence Data; D009928:Organ Specificity; D017386:Sequence Homology, Amino Acid; D013154:Spleen / Q000201:enzymology; D013237:Stereoisomerism; D013379:Substrate Specificity; D013737:Testis / Q000201:enzymology; D014212:Tretinoin / Q000378:metabolism*
D018076:DNA, Complementary; D000077556:Alitretinoin; D014212:Tretinoin; D000429:Alcohol Oxidoreductases; C105846:retinol dehydrogenase 5