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ROLE OF HEAT SHOCK PROTEINS IN PROTEIN FOLDING

热休克蛋白在蛋白质折叠中的作用

基本信息

批准号：
2183071
负责人：
ANTHONY L FINK
金额：
$ 18.73万
依托单位：
UNIVERSITY OF CALIFORNIA SANTA CRUZ
依托单位国家：
美国
项目类别：
财政年份：
1991
资助国家：
美国
起止时间：
1991-01-01 至 1994-12-31
项目状态：
已结题

项目摘要

Mammalian (and other) cells respond to stress by increasing expression of a class of proteins known as heat shock proteins. Most of these proteins are produced constitutively under normal conditions and play essential roles in the life of the cell. Recent studies have shown that the mammalian hsp 70 family of heatshock proteins, and related polypeptide chain binding proteins and molecular chaperones (e.g. GroEL) from other sources, can bind to short peptides, and to nascent polypeptides in cells, and are involved in various aspects of protein assembly, translocation and folding. The relatively large fraction of cell protein, especially under conditions of stress, present as this type of protein, and the potential lethality of mutant forms, indicates their critical importance to cell function. Even though it is clear that proteins fold spontaneously in vitro, it is not clear that this is the case in vivo. In fact, a good case can be made for the need for some form of assistance in folding and assembly in the cell in order to minimize competing side reactions. The goal of the proposed research is to determine the molecular details of how hsp 70 and related polypeptide chain binding proteins facilitate the folding and assembly of substrate proteins. Biophysical characterization of the hsp 70 proteins will provide information on their structure and stability. A major part of the proposal concerns elucidation of the molecular details of the interaction of the hsp 70 with its substrate protein, and the role of ATP in this process. These studies will be carried out under in vitro conditions. The stoichiometry and binding constants for the interactions will be determined. A range of proteins from small monomeric to large oligomeric will be used as substrate proteins. The effect of hsp 70 on the kinetics of protein folding, and the affinity of hsp 70 for intermediates in folding will be determined. The structural and sequence specificity requirements for hsp 70 binding to proteins will be ascertained. From these investigations we expect to be able to provide a detailed molecular description of the function of this important class of cellular agent.

哺乳动物（和其他）细胞通过增加一种蛋白的表达来响应应激。这类蛋白质被称为热休克蛋白。大多数蛋白质是在正常条件下组成产生，并在以下方面发挥重要作用：细胞的寿命。最近的研究表明哺乳动物的hsp70 热休克蛋白家族及相关多肽链结合来自其他来源的蛋白质和分子伴侣（例如GroEL）可以结合短肽和细胞中的新生多肽，在蛋白质组装、移位和折叠的各个方面。的细胞蛋白质的相对大部分，特别是在压力，目前作为这种类型的蛋白质，和潜在的致命性，突变形式，表明它们对细胞功能至关重要。甚至虽然很清楚蛋白质在体外能自发折叠，很明显，这是体内的情况。事实上，一个很好的例子可以证明，在电池中折叠和组装时需要某种形式的辅助，以减少竞争性副反应。建议的目标研究的目的是确定热休克蛋白70及其相关蛋白多肽链结合蛋白促进多肽链的折叠和组装，底物蛋白热休克蛋白70的生物物理特性将提供有关其结构和稳定性的信息。的主要部分该建议涉及阐明的分子细节的热休克蛋白70与其底物蛋白的相互作用，以及ATP的作用在这个过程中。这些研究将在体外进行，条件相互作用的化学计量和结合常数将被确定。一系列蛋白质，从小单体到大寡聚体将用作底物蛋白。热休克蛋白70对人外周血淋巴细胞增殖的影响蛋白质折叠动力学和热休克蛋白70对中间体的亲和力将被确定。结构和序列特异性将确定HSP 70与蛋白质结合的要求。从我们希望这些研究能够提供一个详细的分子描述了这类重要的细胞代理的功能。