Folding And Stability Of Protein Domains and Sub-Domains

描述（由申请人提供）：本提案描述了实验 体外蛋白质折叠的研究。阐明了氨基酸序列 确定结构;即，蛋白质折叠问题是 现代结构生物学最近的两个事态发展使这个问题 特别及时。大规模的测序工作导致了博士的爆炸。 罗利关于蛋白质一级序列的知识。不幸的是 仍然不可能从序列中预测结构。原则上，A 对折叠过程的透彻理解将有助于破译 连接序列和结构的代码。详细了解折叠 这一过程还将有助于合理地修饰蛋白质，以增强所需的功能。 特性.越来越多的人认识到蛋白质的错误折叠和聚集 在一些不同的疾病中发挥作用也引起了人们的关注， 折叠的问题。缺乏结构特异性或在结构中的分解 折叠的协同性可具有严重的生理后果，并且 部分折叠状态在病理生理蛋白质中起重要作用 聚合来本建议侧重于几个实验研究 蛋白质折叠的关键问题：（1）部分蛋白质的结构异质性 蛋白质的折叠状态（2）折叠中协同性的起源（3） 亚结构域在折叠中的作用（4）整个链拓扑结构的作用， 快速折叠中的特异性相互作用。熔化的球状体状态是 在折叠的中间。结合蛋白质切割方法， 对突变体的研究将用于精确定位熔融球的关键区域 α-乳白蛋白状态。现在人们认识到，蛋白质可以聚集在 各种不同类型的部分折叠状态。比较研究 α-乳白蛋白/溶菌酶家族的不同成员的组合将提供 关于不同类型的部分折叠的起源的信息 states.动力学实验将阐明参与的步骤， 从熔化的小球中合作形成自然状态。动力学 用三种小的、快速折叠的螺旋蛋白质进行实验， 关于快速折叠的基本步骤的信息。

DESCRIPTION (provided by applicant): This proposal describes experimental studies of protein folding in vitro. Elucidating how the amino acid sequence determines structure; i.e., the protein folding problem, is a central issue in modern structural biology. Two recent developments have made this problem particularly timely. Massive sequencing efforts have led to an explosion in Dr. Raleigh's knowledge of the primary sequence of proteins. Unfortunately, it is still impossible to predict structure from sequence. In principle, a more thorough understanding of the folding process will aid efforts to decipher the code that links sequence and structure. A detailed understanding of the folding process will also aid efforts to rationally modify proteins to enhance desired properties. The growing realization that protein misfolding and aggregation play a role in a number of different diseases has also focussed attention on the folding problem. A lack of structural specificity or a breakdown in the cooperativity of folding can have severe physiological consequences, and partially folded states play an important role in pathophysiological protein aggregation. This proposal focuses on the experimental investigation of several key issues in protein folding: (1) the structural heterogeneity of partially folded states of proteins (2) the origins of cooperativity in folding (3) the role of subdomains in folding (4) the role of overall chain topology and specific interactions in rapid folding. The molten globule state is a key intermediate in folding. A protein dissection approach in combination with studies of mutants will be used to pinpoint key regions of the molten globule state of alpha-lactalbumin. It is now recognized that proteins can populate a wide range of different types of partially folded states. Comparative studies of different members of the alpha-lactalbumin/lysozyme family will provide information about the origins of the different types of partially folded states. Kinetic experiments will elucidate the steps involved in the cooperative formation of the native state from the molten globule. Kinetic experiments with three small, rapid folding, helical proteins will provide information about the fundamental steps in rapid folding.