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.

由酒精喂养诱导的微粒体酶也是 同样的酶负责血清1，2- 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 乙醇诱导细胞色素P450多克隆抗体 （P450 j，P450 IIE）在兔中制备并用于分离 P450 j的cDNA。 大鼠和小鼠P450 j基因的cDNA序列均经测序证实， 人被分离和表征。 一级氨基酸 通过核苷酸测序法测定了大鼠和人P450 j的序列， P450 j最长cDNA的测序;大鼠P450 j 序列与其他细胞色素P450的序列进行了比较， 确定结构相似性。 用克隆的P450 j cDNA和抗体研究了P450 j基因的表达。 P450 j基因在发育过程中的表达， 诱导物以及病理生理条件。 相结合 cDNA杂交和免疫印迹分析，三种类型的 观察到P450 j基因表达：转录激活 在发育过程中;转录后激活（可能通过 蛋白质稳定化）通过各种诱导剂如乙醇，丙酮， 和吡唑衍生物;以及糖尿病和 饥饿的动物 P450 j在不同组织中的分布及调控 用克隆的P450 j cDNA对组织进行了研究。