UGT2B23, a novel uridine diphosphate-glucuronosyltransferase enzyme expressed in steroid target tissues that conjugates androgen and estrogen metabolites.
Glucuronidation is widely accepted as a mechanism involved in the catabolism and elimination of steroid hormones from the body. However, relatively little is known about the enzymes involved, their specificity for the different steroids, and their site of expression and action. To characterize the pathway of steroid glucuronidation, a novel uridine diphosphate glucuronosyltransferase (UGT) enzyme was cloned and characterized. A 1768-bp complementary DNA, encoding UGT2B23 was isolated from a monkey liver library. Stable expression of UGT2B23 in human HK293 cells and Western blot analysis demonstrated the presence of a 51-kDa protein. The UGT2B23 transferase activity was tested with 62 potential endogenous substrates and was demonstrated to be active on 6 steroids and the bile acid, hyodeoxycholic acid. Kinetic analysis yielded apparent Michaelis constant (Km) values of 0.9, 13.5, 1.6, and 5.7 microM for the conjugation of androsterone (ADT), 3alpha-Diol, estriol, and 4-hydroxyestrone, respectively. RT-PCR analysis revealed that UGT2B23 transcript is expressed in several tissues, including the prostate, mammary gland, epididymis, testis, and ovary. Primary structure analysis shows that UGT2B23 is in the same family of enzymes as the previously characterized monkey isoforms UGT2B9 and UGT2B18, which are active on hydroxyandrogens. The characterization of UGT2B23 as a functional enzyme active on 3alpha-hydroxysteroids, and its expression in extrahepatic tissues, indicate that it may potentially play an important role in estrogen and androgen catabolism in peripheral steroid target tissues.
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Laboratory of Molecular Endocrinology, Centre Hospitalier de L'Université Laval Research Center, Laval University, Québec, Canada.
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D000595:Amino Acid Sequence; D000728:Androgens / Q000378:metabolism*; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002460:Cell Line; D018076:DNA, Complementary / Q000737:chemistry / Q000302:isolation & purification; D004967:Estrogens / Q000378:metabolism*; D005260:Female; D015870:Gene Expression; D020719:Glucuronides / Q000378:metabolism; D014453:Glucuronosyltransferase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D000882:Haplorhini; D006801:Humans; D007700:Kinetics; D008297:Male; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000032:analysis; D016415:Sequence Alignment; D014018:Tissue Distribution; D014162:Transfection
D000728:Androgens; D018076:DNA, Complementary; D004967:Estrogens; D020719:Glucuronides; D012333:RNA, Messenger; D014453:Glucuronosyltransferase