Identification and analysis of drug-responsive expression of UDP-glucuronosyltransferase family 1 (UGT1) isozyme in rat hepatic microsomes using anti-peptide antibodies.
Identification and analysis of drug-responsive expression of UDP-glucuronosyltransferase family 1 (UGT1) isozyme in rat hepatic microsomes using anti-peptide antibodies.
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使用抗肽抗体鉴定和分析大鼠肝微粒体中 UDP-葡萄糖醛酸基转移酶家族 1 (UGT1) 同工酶的药物反应性表达。
DOI：
10.1006/abbi.1995.0039
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发表时间：
1995
影响因子：
3.9
通讯作者：
T. Iyanagi
中科院分区：
文献类型：
--
作者：
Shinichi Ikushiro;Y. Emi;T. Iyanagi
利用抗肽抗体检测了大鼠肝脏udp -葡萄糖醛酸糖基转移酶家族1 （UGT1）同工酶的表达，这些抗体针对所有同工酶的保守羧基末端部分和酚簇（UGT1A）和胆红素簇（UGT1B）的每个同工酶的可变氨基末端部分。在大鼠肝微粒体中表达的同工酶中，胆红素簇的UGT1B1 （54 kDa）为主要形式，UGT1A1 （53 kDa）、UGT1B2 （56 kDa）和UGT1B5 （57 kDa）为次要形式。利用二维十二烷基硫酸钠凝胶电泳和免疫印迹相结合的方法，发现Gunn大鼠肝微粒体同时缺乏所有同工酶。分析不同药物作为诱导剂对UGT1各同工酶表达的影响。3-甲基胆蒽处理大鼠的酚簇同工酶UGT1A1和UGT1A2均被显著诱导。氯贝特和地塞米松可诱导大鼠肝微粒体中UGT1B1的表达和对胆红素的糖醛酸化活性增加2 ~ 3倍。另一方面，对UGT1B2和UGT1B5表达的调控与UGT1B1不同。这些结果首次用同工酶特异性抗体清楚地显示了UGT1各同工酶的药物反应性表达。
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.