Protein Conformation and Noncovalent Interactions

描述（由申请人提供）：本研究的目的是研究确定蛋白质结构、测量蛋白质-蛋白质结构和结合的新方法，以及使用质谱法分离和鉴定不同蛋白质构象异构体。将进行溶液相和气相研究。从这两个阶段的结构或结合相互作用的差异，可以确定有关溶剂如何影响蛋白质构象和蛋白质之间的特定分子间相互作用的信息。这些信息可能会增强确定蛋白质结构和折叠的计算方法以及药物发现的质谱方法。一种灵敏、高通量的蛋白质构象测定方法可以大大提高研究人员发现蛋白质功能和鉴定新结构药物的能力。将研究非共价复合物的串联质谱实验。这些研究可以提供其他方法难以获得或无法获得的结构信息。具体目标包括：1）开发一种潜在灵敏和快速的方法，用于使用具有电子捕获解离的溶液相H/D交换来确定蛋白质构象，以识别具有单个氨基酸分辨率的交换位点，2）评估蛋白质-蛋白质复合物中的溶液相和气相结合相互作用，以及3）研究高-场不对称波形离子迁移谱是一种快速、灵敏的蛋白质构象分析方法。人们希望，这些研究将提供一个坚实的基础相关的结构信息的生物聚合物和非共价复合物从气相实验回到本体溶液中的离子的结构。这项研究旨在开发新的方法来快速确定蛋白质的折叠结构，它们如何与其他蛋白质相互作用，以及周围的溶剂分子如何影响这些相互作用。这些研究可以提供重要的新信息，这些信息可用于理解蛋白质错误折叠的疾病，包括阿尔茨海默病、囊性纤维化、海绵状脑病（例如，疯牛病或克雅氏病），甚至一些癌症。此外，蛋白质-蛋白质相互作用的研究可以潜在地提供一种更快和更通用的方法，可以显着改善用于破坏经常与人类疾病相关的异常复合物的新药的发现。

DESCRIPTION (provided by applicant): The goals of this research are to investigate new methods to determine protein structure, measure protein-protein structure and binding, and separate and identify different protein conformers using mass spectrometry methods. Both solution-phase and gas-phase studies will be performed. From differences in structure or binding interactions in these two phases, information about how solvent influences both protein conformation and specific intermolecular interactions between proteins can be determined. This information could potentially enhance computational methods for determining protein structure and folding and for mass spectrometry methods for drug discovery. A sensitive, high throughout method for determining protein conformation could greatly improve researchers' ability to discover functions of proteins and identify new structure based medicines. Tandem mass spectrometry experiments of noncovalent complexes will be investigated. These studies can provide structural information that is difficult or not obtainable by other methods. Specific aims include 1) develop a potentially sensitive and rapid method for determining protein conformation using solution-phase H/D exchange with electron capture dissociation for identifying exchange sites with individual amino acid resolution, 2) evaluate both solution-phase and gas-phase binding interactions in a protein-protein complex and 3) investigate high-field asymmetric waveform ion mobility spectroscopy as a rapid and sensitivity method for protein conformational analysis. It is hoped that these studies will provide a firm basis for relating structural information of biopolymers and noncovalent complexes determined from gas-phase experiments back to the structures of the ions in bulk solution. This research is aimed at developing new methods for rapidly determining the folded structure of proteins, how they interact with other proteins, and how surrounding solvent molecules can effect these interactions. These studies can provide important new information that can be useful for understanding diseases in which proteins misfold, including Alzheimer's disease, cystic fibrosis, spongiform encephalopathies (e.g., Mad Cow or Creutzfeldt Jakob disease), and even some cancers. In addition, the studies of protein-protein interactions can potentially provide a faster and more general method that could significantly improve the discovery of new drugs for disrupting aberrant complexes that are frequently associated with human disease.