Protein Conformation and Noncovalent Interactions

本研究的目的是探索新的方法来确定蛋白质结构，测量 蛋白质-蛋白质结构和结合，并分离和鉴定不同的蛋白质构象使用质量 光谱法将进行溶液相和气相研究。的差异所 这两个阶段的结构或结合相互作用，关于溶剂如何影响蛋白质 可以确定蛋白质之间的构象和特异性分子间相互作用。这些信息 可以潜在地增强确定蛋白质结构和折叠以及质量的计算方法 用于药物发现的光谱分析方法。一种灵敏、高通量的蛋白质测定方法 构象可以大大提高研究人员发现蛋白质功能和识别新的 基于结构的药物非共价复合物的串联质谱实验将在 研究了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。 方法.具体目标包括：1）开发一种潜在的敏感和快速的蛋白质测定方法 使用具有电子捕获解离的溶液相H/D交换来鉴定交换的构象 具有单个氨基酸分辨率的位点，2）评估溶液相和气相结合 蛋白质-蛋白质复合物中的相互作用和3）研究高场不对称波形离子迁移率 光谱法是一种快速、灵敏的蛋白质构象分析方法。希望这些 研究将为生物聚合物和非共价复合物的结构信息的关联提供坚实的基础 从气相实验确定回到本体溶液中离子的结构。 本研究的目的是发展新的方法，快速确定折叠结构， 蛋白质，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 交互.这些研究可以提供重要的新信息， 蛋白质错误折叠的疾病，包括阿尔茨海默氏病、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。此外该 蛋白质-蛋白质相互作用的研究可能提供一种更快和更通用的方法， 显着提高了发现新的药物，破坏异常复合物， 与人类疾病有关。

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.