IMMUNOPHARMACOLOGICAL IDENTIFICATION AND REGULATION OF CYTOCHROME P-450

本研究项目的目的是了解该系统的功能和 细胞色素P-450不同形态的生理调节 以免疫学为基础的方法。正在采取的方法是 产生抗肽抗体，这允许产生抗体 它们是单一的P-450酶所特有的，而不是特定的P-450家族。 P-450特异性抗体的发展使我们能够更准确地 确定P-450酶在选定的代谢中的作用 底物。目前的研究主要集中在多克隆的发展上。 以及针对P450 IA2特有序列的单抗。这架P-450 酶是杂环胺类致癌物代谢的关键酶。 在P-450IA2中已经发现了几个独特的区域。目前，有 目前还没有能识别P-450IA2的抗体。另一个 这项工作的一个方面是阐明了P-450在多大程度上 IA2参与了乙酰苯胺的代谢。使用单抗 1-7-1，识别P-450 LAL和IA2，以及cDNAs表达的小鼠 IAI和小鼠以及人的IA2，我们能够证明IA2是 乙酰苯胺向其代谢的主要P-450形式 羟化代谢物。P450的IAI、IIB1和IIB2不代谢 乙酰苯乙胺。单抗1-7-1抑制了80%的乙酰苯胺羟基酶 小鼠和人都表达了IA2的活性，尽管是24倍以上 需要抗体才能最大限度地抑制人IA2 达到了小鼠IA2所需的水平。单抗1-7-1抑制乙酰苯胺 3-甲基胆蒽诱导的肝微粒体羟基酶活性 也有80%。

The goal of this research project is to understand the function and physiological regulation of the various forms of cytochrome P-450 using immunologically-based methodologies. The approach being undertaken is to generate anti-peptide antibodies, which permit the creation of antibodies which are specific for single P-450 enzyme and not a given P-450 family. The development of P-450 specific antibodies permits us to more precisely define the role of that P-450 enzyme in the metabolism of selected substrates. Current research is focused on the development of polyclonal and monoclonal antibodies against sequences unique to P450 IA2. This P-450 enzyme is a key enzyme in the metabolism of heterocyclic amine carcinogens. Several unique regions have been identified in P-450 IA2. Currently, there is no available antibody which specifically recognizes P-450 IA2. Another aspect to this work has been the elucidation of the extent to which P-450 IA2 is involved in the metabolism of acetanilide. Using monoclonal antibody 1-7-1, which recognizes both P-450 lAl and IA2, and cDNA-expressed mouse IAI and mouse and human IA2, we were able to demonstrate that IA2 is the major P-450 form responsible for the metabolism of acetanilide to its hydroxylated metabolites. P450s IAI, IIB1 and IIB2 did not metabolize acetanilide. MAb 1-7-1 inhibited 80% of the acetanilide hydroxylase activity of both mouse and human cDNA expressed IA2, though 24-fold more antibody was needed to achieve maximal inhibition of human IA2 as compared to that needed for mouse IA2. MAb 1-7-1 inhibited the acetanilide hydroxylase activity of 3-methylcholanthrene-induced liver microsomes by 80% also.