STRUCTURE AND REGULATION OF ETHANOL-INDUCIBLE CYTOCHROME P450 GENE

乙醇诱导细胞色素P450基因的结构和调控

• 批准号: 3821232
• 负责人: B J SONG
• 金额: --
• 依托单位: NATIONAL INSTITUTE ON ALCOHOL ABUSE AND ALCOHOLISM
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3821232
• 关键词: antibody formation; complementary DNA; cytochrome P450; ethanol; fasting; gene expression; genetic mapping; genetic regulation; genetic transcription; human tissue; messenger RNA; molecular cloning; nucleic acid sequence; pyrazoles

The microsomal enzyme induced by feeding of alcohol is also the same enzyme responsible for the elevation of serum 1,2- propanediol found in the serum of alcoholics (Koop, D.R., Casazza, J.P.: J Biol Chem 260: 13607-13612, 1985). Specific polyclonal antibody against ethanol-inducible cytochrome P450 (P450j, P450 IIE) was prepared in rabbit and utilized to isolate cDNA for P450j. The cDNAs encoding P450j of both rat and human were isolated and characterized. The primary amino acid sequences of rat and human P450j were determined by nucleotide sequencing of the longest cDNAs for P450j; The rat P450j sequence was compared to those of other cytochrome P450s to determine the structural similarity. Cloned P450j cDNA and antibodies were used to study the expression of P450j gene during development and by various inducers as well as pathophysiological conditions. By combination of cDNA hybridization and immunoblot analyses, three types of P450j gene expression were observed: transcriptional activation during development; post-transcriptional activation (probably via protein stabilization) by various inducers such as ethanol, acetone, and pyrazole derivatives; and mRNA stabilization in diabetes and starved animals. Distribution and regulation of P450j in different tissues were also studied by using cloned P450j cDNA.

通过饮酒诱导的微粒体酶也是 相同的酶负责血清1,2-的升高 在酗酒者的血清中发现的丙二醇（Koop，D.R。， Casazza，J.P。：J Biol Chem 260：13607-13612，1985）。 具体的 抗乙醇可诱导的细胞色素P450的多克隆抗体 （P450J，P450 IIE）是在兔子中制备的，并用于分离 P450J的cDNA。 编码大鼠和大鼠和 人是孤立和表征的。 初级氨基酸 大鼠和人P450J的序列通过核苷酸测定 P450J最长cDNA的测序； 大鼠P450J 将序列与其他细胞色素P450的序列进行比较 确定结构相似性。 克隆的P450J cDNA和抗体用于研究 P450J基因在发育过程中和各种表达 诱导者以及病理生理状况。 结合 cDNA杂交和免疫印迹分析，三种类型 观察到P450J基因表达：转录激活 在发展期间；转录后激活（可能是通过 蛋白质稳定）由各种诱导剂，例如乙醇，丙酮， 和吡唑衍生物；糖尿病和mRNA稳定 饥饿的动物。 P450J在不同的分布和调节 还使用克隆的P450J cDNA研究了组织。