PROTEIN FOLDING IN THE EUKARYOTIC CYTOSOL

描述：这项建议的长期目标是了解 真核细胞中蛋白质折叠的生物化学和细胞生物学。这个 一般的策略将是专注于折叠两种模式 蛋白质、肌动蛋白和荧光素酶。拟议的研究将集中在折叠方面 在多肽合成过程中发生在核糖体上的事件 将研究分子伴侣在折叠过程中的作用。这个 理解蛋白质折叠的概念框架 翻译源于申请人的早期作品，这表明 新翻译的多肽被引导到它们的最终 通过一个连续的和高度偶联的伴侣途径进行构象。 这一途径结合了伴侣作用的两个不同原则，即 Hsp70/Hsp40蛋白通过稳定延伸的多肽而起作用，以及 守护结构凭借其环形结构创建了一个 有利于多肽链折叠的环境。 该提案的目标是调查 通过以下方式折叠新合成的蛋白质：1)定义 与新生的链相互作用的伴侣分子，如多肽长度 增加，2)评估单个伴侣蛋白的需求量 新合成蛋白质的折叠并决定是否折叠 与伴侣复合体相关发生，3)调查角色 的翻译装置和可能的新颖组件 招募新成立的连锁店的监护人，以及4)审查 完整分子伴侣与新翻译多肽的相互作用 哺乳动物细胞。

DESCRIPTION: The long term goal of this proposal is to understand the biochemistry and cell biology of protein folding in eukaryotic cells. The general strategy will be to concentrate on the folding of two model proteins, actin and luciferase. The proposed research will focus on folding events as they occur at the ribosome during synthesis of a polypeptide and will examine the role of molecular chaperones in the folding process. The conceptual framework to understand the folding of proteins during translation originates from the applicant's earlier work, which indicates that the newly translated polypeptides are guided to their final conformation through a sequential and highly coupled chaperone pathway. This pathway combines two distinct principles of chaperone action, namely the Hsp70/Hsp40 proteins that act by stabilizing extended polypeptides, and the chaperonin TRiC, which by virtue of its ring-like structure creates an environment that is favorable for polypeptide chain folding. The objective of the proposal is to investigate critical aspects of the folding of newly synthesized proteins by: 1) defining the complement of chaperones that interact with nascent chains as the polypeptide length increases, 2) assessing the requirement of individual chaperone proteins in the folding of newly synthesized proteins and determine whether folding occurs in association with the chaperone complex, 3) investigating the role of the translational apparatus and possible novel components in the recruitment of chaperones to the nascent chains, and 4) examining the interaction of chaperones with newly translated polypeptides in intact mammalian cells.