Identification and analysis of drug-responsive expression of UDP-glucuronosyltransferase family 1 (UGT1) isozyme in rat hepatic microsomes using anti-peptide antibodies.
Expression of rat hepatic UDP-glucuronosyltransferase family 1 (UGT1) isozymes has been examined using anti-peptide antibodies raised against a conserved carboxyl-terminal portion of all isozymes and variable amino-terminal portions of each isozyme of the phenol cluster (UGT1A) and bilirubin cluster (UGT1B). Among the isozymes expressed in rat hepatic microsomes, UGT1B1 (54 kDa) of bilirubin cluster was found to be a major form and minor forms were identified as UGT1A1 (53 kDa), UGT1B2 (56 kDa), and UGT1B5 (57 kDa). Using a combination of 2D sodium dodecyl sulfate gel electrophoresis and immunoblotting, all the isozymes were found to be simultaneously lacked in Gunn rat hepatic microsomes. The effects of various drugs as inducer on the expression of each UGT1 isozyme were analyzed. The UGT1A1 and UGT1A2 of the phenol cluster isozymes were significantly induced in 3-methylcholanthrene-treated rats. The expression of UGT1B1 and the glucuronidation activity toward bilirubin in rat hepatic microsomes were induced two- to threefold by clofibrate and dexamethasone administration. On the other hand, the regulation of UGT1B2 and UGT1B5 expression was different from that of UGT1B1. These results clearly show the drug-responsive expression of each UGT1 isozyme using isozyme-specific antibodies for the first time.
Ikushiro
S
S
Department of Life Science, Faculty of Science, Himeji Institute of Technology, Hyogo, Japan.
Emi
Y
Y
Iyanagi
T
T
D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D001663:Bilirubin / Q000494:pharmacology; D002994:Clofibrate / Q000494:pharmacology; D003907:Dexamethasone / Q000494:pharmacology; D004790:Enzyme Induction; D014453:Glucuronosyltransferase / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification; D007527:Isoenzymes / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology*; D008969:Molecular Sequence Data; D009596:Nitrophenols / Q000494:pharmacology; D009842:Oligopeptides / Q000138:chemical synthesis / Q000276:immunology; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000276:immunology*; D010600:Pharmacology; D051381:Rats; D011911:Rats, Gunn; D017208:Rats, Wistar
D007527:Isoenzymes; D009596:Nitrophenols; D009842:Oligopeptides; D010446:Peptide Fragments; D003907:Dexamethasone; D014453:Glucuronosyltransferase; D002994:Clofibrate; D001663:Bilirubin; C024836:4-nitrophenol