PROTEIN FOLDING IN THE EUKARYOTIC CYTOSOL

描述：本提案的长期目标是了解 真核细胞中蛋白质折叠的生物化学和细胞生物学。 的 总的战略将集中在两个模式的折叠 蛋白质、肌动蛋白和荧光素酶。 拟议的研究将集中在折叠 在多肽合成期间在核糖体发生的事件， 将研究分子伴侣在折叠过程中的作用。 的 理解蛋白质折叠的概念框架 翻译来源于申请人的早期工作，这表明 新翻译的多肽被引导到它们的最终 通过连续的和高度偶联的伴侣蛋白途径的构象。 该途径结合了分子伴侣作用的两个不同原则，即 通过稳定延伸多肽起作用的Hsp 70/Hsp 40蛋白，和 伴侣蛋白TRiC，其凭借其环状结构产生一种 这是有利于多肽链折叠的环境。 该提案的目的是调查 新合成的蛋白质的折叠：1）定义 与新生链相互作用的分子伴侣， 增加，2）评估个体伴侣蛋白的需求， 新合成的蛋白质的折叠，并确定折叠是否 发生在与伴侣复合物，3）调查的作用 的平移装置和可能的新的组成部分， 招募分子伴侣的新生链，和4）检查 分子伴侣与完整细胞中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.