REGULATION AND FUCTION OF P450 3A GENE FAMILY ISOZYMES

描述：这是竞争延续的第一个版本。 的 工作分为四个具体目标：SA1。 生产和净化 针对大鼠P450 3A1、3A2和3Ax的Mab试剂组合，以及 相关的人P450 3A3、3A4、3A5和3A7。 建议成员 所需的产物组在ELISA测定，蛋白质印迹， 对微粒体提取物中P450活性抑制， 对于每个P450，将发现与不同表位结合的抗体， 促进P450同源物的种间比较，用于膜拓扑学 免疫细胞化学分析。 SA 2. 为了研究年龄和性别 在mRNA和蛋白水平上对大鼠3A1和3A2的特异性调节。 SA 3. 为了克隆和表征新的3A同位素（3Ax）的cDNA， 最近由主要研究者确定，并研究其 表达作为年龄、性别和组织类型的函数，如SA2中所述。 SA 4. 为了使用在SA 1中获得的抑制性单克隆抗体，选择性地抑制 P450在大鼠和人微粒体提取物中的表达，从而建立 各种同位素在选定底物代谢中的作用 (CsA孕酮等）。

DESCRIPTION: This is the first revision of a competitive continuation. The work is divided into four specific aims: SA1. To produce and purify a battery of Mab reagents directed against rat P450 3A1, 3A2 and 3Ax, and the related human P450s 3A3, 3A4, 3A5 and 3A7. It is proposed that members of the desired product set will be competent in ELISA assays, western blotting, inhibitory towards P450 activity in microsomal extracts, and that several antibodies binding to distinct epitopes will be found for each P450, to facilitate interspecies comparison of P450 homologues, for membrane topology analysis and immunocytochemical analysis. SA2. To study the age and sex specific regulation of 3A1 and 3A2 in the rat at the mRNA and protein level. SA3. To clone and characterize the cDNA for a novel 3A isotope (3Ax), recently identified by the principal investigator, and to study its expression as a function of age, sex and tissue type, as described in SA2. SA4. To use the inhibitory Mab s obtained in SA1, to selectively inhibit P450 expression in rat and human microsomal extracts, so as to establish the function of the various isotopes in the metabolism of selected substrates (CsA, progesterone, and others).