REGULATED DEGRADATION OF ETHANOL INDUCIBLE P450 2E1

乙醇诱导 P450 2E1 的调节降解

• 批准号: 2748419
• 负责人: JIANYA HUAN
• 金额: $ 3.15万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-08-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2748419
• 关键词: chimeric proteins; cytochrome P450; drug metabolism; enzyme inhibitors; enzyme structure; ethanol; protein degradation; protein structure function; proteolysis; site directed mutagenesis

The long term goal of this proposal is to define the proteolytic pathway which degrades CYP2El and to identify those regions of the protein that are important for targeting the enzyme for degradation. The results of this proposal will lead a better understanding in the role of structural motifs in the degradation of CYP2El. The goal of this project will be accomplished through the direct determination of the steady state half- life of the native enzyme, amino- and carboxyl-terminal deleted mutants and chimeric proteins in which regions of the longer half-life CYP2Bl are combined with those of CYP2El. Once a region is defined through analysis of chimeric enzymes, specific site directed mutants will be prepared in order to identify residues which may be important in the determination of the degradative path of the enzyme. In all cases, half-life studies will be coupled with the use of selective inhibitors of key intracellular proteolytic enzymes in order to determine whether chimeric enzymes follow the same degradative path as the native protein. The susceptibility of the various constructs to proteolysis i,i vitro will also be determined.

该提议的长期目标是定义蛋白水解途径 降解CYP2EL并确定蛋白质的那些区域 对于靶向降解酶很重要。结果 该提议将有助于更好地理解结构的作用 CYP2EL降解中的主题。这个项目的目标将是 通过直接确定稳态一半来完成 天然酶，氨基和羧基末端删除突变体的寿命 和嵌合蛋白，其中更长的半衰期CYP2BL的区域为 与CYP2EL的那些结合在一起。一旦通过分析定义了一个区域 在嵌合酶中，将准备特定位置的定向突变体 为了识别可能在确定的残留物 酶的降解路径。在所有情况下，半衰期研究将 与关键细胞内的选择性抑制剂的使用相结合 蛋白水解酶为了确定是否遵循嵌合酶 与天然蛋白质相同的降解路径。的敏感性 蛋白水解I的各种构建体也将确定体外。