CHAPERONES AND THE BIOGENESIS OF MEMBRANE PROTEINS

伴侣分子和膜蛋白的生物发生

• 批准号: 6474958
• 负责人: DOUGLAS M CYR
• 金额: $ 14.51万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-01-01 至 2002-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6474958
• 关键词: Saccharomyces cerevisiae; cell free system; chloride channels; endoplasmic reticulum; membrane biogenesis; membrane proteins; molecular chaperones; protein folding; protein transport; stress proteins

Although much work in recent years has been focused on the role of lumenal ER chaperones in the folding and assembly of secretory proteins, many of these proteins also possess rather long cytosolic portions. It is assumed that they also require the action of chaperones, but this has not been investigated in any detail. Members of the ABC protein family have very long cytosolic domains that encompass a major portion of the protein. It is well recognized that dnaJ homologues interact with dnaK homologues (hsp70s) to promote the folding of substrate proteins. Cytosolic, ER localized, dnaJ homologues (hsp40s) have been identified in yeast and mammals. It is the hypothesis of this proposal that YDJ-1 (yeast) and HDJ- 1 and -2 (mammalian) collaborate with cytosolic hsp70 to fold the cytosolic tail of ER membrane proteins. In the first aim, the P.I. will use genetic and biochemical techniques to examine the role of YDJ-1 and hsp70 and Ste6 folding in S. cerevisiae. In the second aim, he will develop a cell free system to characterize reaction intermediates in the pathway for CFTR folding. In the third aim, purified components will be employed to determine the molecular mechanism by which hsp40/hsp70 chaperone pairs facilitate the folding of soluble cytosolic domains on membrane proteins.

虽然近年来的许多工作集中在Lumenal的作用上 ER伴侣在分泌蛋白的折叠和组装中，许多 这些蛋白质还具有相当长的胞质部分。假设 他们还需要伴侣的作用，但这不是 详细调查。 ABC蛋白家族的成员非常 涵盖蛋白质主要部分的长胞质结构域。它 众所周知，DNAJ同源物与DNAK同源物相互作用 （HSP70S）促进底物蛋白的折叠。胞质，ER 在酵母中已经鉴定出局部的DNAJ同源物（HSP40） 哺乳动物。 YDJ-1（酵母）和HDJ-是该提议的假设 1和-2（哺乳动物）与胞质HSP70合作折叠 ER膜蛋白的胞质尾巴。在第一个目标中，P.I.将要 使用遗传和生化技术来检查YDJ-1和 Hsp70和Ste6在酿酒酵母中折叠。在第二个目标中，他将 开发无细胞系统以表征反应中间体 CFTR折叠的途径。在第三个目标中，纯化的组件将是 用于确定hsp40/hsp70的分子机制 伴侣对促进可溶性胞质结构域的折叠 膜蛋白。