IMMUNOPURIFICATION AND CHARACTERIZATION OF CYTOCHROME P-450

细胞色素P-450代谢多种化合物， 包括外源性物质，如药物和致癌物质， 内源性化合物如类固醇。 本项目的重点 是结构-功能关系的表征， 调节这种酶的多种形式。 单克隆 抗大鼠P-450的抗体（MAb）是这些研究中的工具。 A 3- 免疫纯化甲基胆蒽（MC）诱导的P-450 从老鼠的肝脏和肺中提取的 以氨基酸为基础 序列分析、肽图谱和分子量， 肝和肺的P-450无法区分。 使用单克隆抗体 乙醇诱导的大鼠肝P-450，P-450已从 大鼠和人的肝脏。 它们的一级结构不同， 通过不同的氨基末端序列和肽证明 地图 用免疫组织化学方法检测P-450的发育调节。 研究P-450依赖的睾酮代谢在3-和24- 一个月的老鼠 环羟基化模式，以及 肝脏组成性P-450随年龄变化。 虽然P-450的活动 通常随着年龄的增长而下降，7 α-羟化酶和 负责这一活动的P-450表格的相应数量增加 随年龄

The cytochromes P-450 metabolize a wide array of compounds, including xenobiotics such as drugs and carcinogens, and endogenous compounds such as steroids. The focus of this project is the characterization of structure-function relationships and regulation of the multiple forms of this enzyme. Monoclonal antibodies (MAbs) to rat P-450s are a tool in these studies. A 3- methylcholanthrene (MC)-inducible P-450 was immunopurified from the livers and lungs of rats. On the basis of amino acid sequence analysis, peptide mapping, and molecular weight, the liver and lung P-450s were indistinguishable. Using a MAb to ethanol-inducible rat liver P-450, a P-450 has been purified from both rat and human liver. These differed in primary structure as evidenced by different amino terminal sequences and peptide maps. Developmental regulation of P-450 was examined by studying P-450-dependent testosterone metabolism in 3- and 24- month old rats. Ring hydroxylation patterns, as well as content of liver constitutive P-450s, varied with age. While P-450 activities generally declined with age, the 7 alpha-hydroxylase and corresponding P-450 form responsible for this activity increased with age.