UDP-GLUCURONOSYLTRANSFERASES

治疗有用的药物和内生菌的葡萄糖醛酸化作用 类固醇和胆红素调节药物的作用时间和毒性 这些物质，尤其是在人类体内。一个微粒体UDP家族-- 葡萄糖醛酸基转移酶(UDPGTs)催化这些反应，并在某些情况下 4例中，人类肝脏中存在独特的UDPGT。这项提议的目的是 揭示了UDPGTS介导的功能和结构特性 吗啡、叔胺的3-0和6-0-葡萄糖醛酸化反应 吗啡(如三苯那胺、阿米替林)。具体地说，吗啡 将从大鼠和人肝微粒体中纯化UDPGT以获得均一 并与它们的底物专一性、NH2末端进行比较 氨基酸序列和活性部位肽氨基酸序列。Cdna 将使用聚合酶克隆大鼠和人肝脏的UDPGT 与寡核苷酸的链式反应技术的合成将是 基于氨基酸序列知识。特定活性部位的光亲和力 使用[~3H]-氟硝西潘(FNZ)标记将有助于实现 这些目标的数量。活性和FNZ标记的吗啡UDPGT将是 使用FractoGel、层析聚焦和亲和纯化至均一 层析法。人肝叔胺UDPGT也将被研究 利用光亲和标记，选择抗体免疫识别， 层析分离、氨基末端氨基酸测序及cDNA 克隆技术。与性别有关或药物导致的人类数量增加 肝脏雌三醇UDPGT也将被研究。这项研究将提供 能够预测潜在药物的信息 UDPGT的相互作用、研究方法或可能的人类多态， 了解葡萄糖糖醛酸化作用对新陈代谢的调节。

Glucuronidation of therapeutically useful drugs and endobiotics such as steroids and bilirubin regulates the duration of action and toxicity of these substances, especially in humans. A family of microsomal UDP- glucuronosyltransferases (UDPGTs) catalyze these reactions and, in certain cases, unique UDPGTs exist in human liver. The objective of this proposal is to reveal the functional and structural properties of UDPGTs mediating the 3-0 and 6-0-glucuronidation of morphine, tertiary amines other than morphine (e.g., tripelennamine, amitryptyline). Specifically, morphine UDPGTs will be purified to homogeneity from rat and human liver microsomes and compared with respect to their substrate specificity, NH2-terminal amino acid sequences and active site peptide amino acid sequences. cDNA cloning of rat and human liver UDPGTs will be performed using polymerase chain reaction techniques with oligonucleotides whose synthesis will be based on amino acid sequence knowledge. Specific active site photoaffinity labeling using [3H]-flunitrazepam (FNZ) will be useful in accomplishing a number of these aims. Active and FNZ-labeled morphine UDPGT will be purified to homogeneity using Fractogel, chromatofocusing and affinity chromatography. Human liver tertiary amine UDPGT will also be studied using photoaffinity labeling, selected antibody immunorecognition, chromatographic separations, NH2-terminal amino acid sequencing and cDNA cloning techniques. Gender-related or drug-induced increases in human liver estriol UDPGT will also be studied. This research will provide information which will allow for predictions of potential drug-drug interaction, methods for study or possible human polymorphisms of UDPGTs, and an understanding of the regulation of metabolism by glucuronidation.