Spectroscopic Study of Protein Folding Dynamics

描述（由申请人提供）：蛋白质通过其线性多肽链的特异性折叠获得其独特功能。错误折叠导致许多疾病，如囊性纤维化和各种神经退行性疾病。虽然已经做了大量的工作来研究蛋白质的二级和三级结构是如何形成的，并且用于研究蛋白质折叠的实验和理论技术都在不断地变得更加完善，但是仍然无法实现对蛋白质折叠的定量和预测性理解。仍然有许多基本的问题，如：如何与周围环境的特定和非特异性相互作用确定的折叠途径，能量景观的粗糙度，以及热和动力学可访问的构象基板？在拥挤的环境和/或膜中，特定的构象和折叠事件发生的时间范围是多少？解决这些问题提出了需要进一步的研究与时间分辨光谱技术，可以提供必要的时间分辨率和结构灵敏度。因此，拟议的研究的主要目标是开发新的光谱方法和新的构象探针，可用于产生详细的结构解释的瞬态折叠物种和它们的动态在很宽的时间尺度。一套全面的实验计划，以获得新的见解，了解蛋白质折叠问题的各个方面，以及膜蛋白的结构-动力学-功能的关系。技术目标是：（a）开发新的光谱探针和方法用于蛋白质折叠和结合研究;（B）研究大分子拥挤和限制如何影响蛋白质折叠和聚集的动力学;（c）研究膜蛋白质折叠的早期动力学事件;（e）探测膜蛋白质功能的构象动力学。实现这些特定目标不仅会为各种生物学应用带来新的实验方法，而且还会带来新的发现，有助于提高我们对蛋白质折叠中许多基本问题的理解，从而为该领域做出实际和直接的贡献。 公共卫生相关性：蛋白质通过其线性多肽链的特异性折叠获得其独特功能。错误折叠导致许多疾病，如囊性纤维化和各种神经退行性疾病。实现这些具体目标不仅会导致新的折叠和结合研究的实验方法，而且会对蛋白质折叠中的许多基本问题的理解产生新的见解，从而对该领域做出实际和直接的贡献，并对公共卫生具有根本的重要性。

DESCRIPTION (provided by applicant): Proteins acquire their unique functions through specific folding of their linear polypeptide chains. Misfolding results in numerous diseases, such as cystic fibrosis and various neurodegenerative disorders. Although a great deal of work has been done to investigate how the secondary and tertiary structures of proteins are formed and both experimental and theoretical techniques for studying protein folding are continually becoming more refined, a quantitative and predictive understanding of protein folding is still not attainable. There are still many fundamental questions, such as: How do specific and nonspecific interactions with the surroundings determine the folding pathways, the roughness of the energy landscape, and the thermally and kinetically accessible conformational substrates? On what range of timescales do particular conformational and folding events occur in crowded environments and/or membranes? Addressing these questions presents the need for further studies with time-resolved spectroscopic techniques that can provide the necessary time resolution and structural sensitivities. The principal objective of the proposed research is therefore to develop new spectroscopic methods and new conformational probes that can be used to generate detailed structural interpretations of the transient folding species and their dynamics over a wide range of timescales. A comprehensive set of experiments are planned to gain new insight into the understanding of various aspects of the protein folding problem, as well as the structure-dynamics-function relationship of membrane proteins. The technical goals are to: (a) develop new spectroscopic probes and methods for protein folding and binding studies; (b) investigate how macromolecular crowding and confinement affect the kinetics of protein folding and aggregation; (c) study the early kinetic events in membrane protein folding; (e) probe the conformational dynamics underlying membrane protein function. Achieving these specific aims should result in not only new experimental methods for various biological applications, but also new findings that would help improve our understanding of many fundamental issues in protein folding, thus making practical and direct contributions to the field. PUBLIC HEALTH RELEVANCE: Proteins acquire their unique functions through specific folding of their linear polypeptide chains. Misfolding results in numerous diseases, such as cystic fibrosis and various neurodegenerative disorders. Achieving these specific aims should result in not only new experimental methods for folding and binding studies, but also new insights into the understanding of many fundamental issues in protein folding, thus making practical and direct contributions to the field and having fundamental importance for public health.