Protein Conformation and Noncovalent Interactions

描述（由申请人提供）：本研究的目的是探索利用质谱法测定蛋白质结构、测量蛋白质-蛋白质结构和结合以及分离和鉴定不同蛋白质构象的新方法。溶液相和气相研究都将进行。从这两个阶段的结构差异或结合相互作用中，可以确定溶剂如何影响蛋白质构象和蛋白质之间特定的分子间相互作用的信息。这一信息可能潜在地增强用于确定蛋白质结构和折叠的计算方法以及用于药物发现的质谱方法。一种灵敏、高通量的测定蛋白质构象的方法可以极大地提高研究人员发现蛋白质功能和鉴定基于新结构的药物的能力。将研究非共价配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)开发一种潜在的敏感和快速的方法来确定蛋白质构象，使用溶液相H/D交换和电子捕获解离来识别单个氨基酸分辨率的交换位点；2)评估蛋白质复合物中的溶液相和气相结合相互作用；3)研究高场不对称波形离子迁移谱作为蛋白质构象分析的快速和敏感方法。希望这些研究将为从气相实验中确定的生物聚合物和非共价配合物的结构信息与体溶液中离子的结构联系起来提供坚实的基础。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，有助于了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（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能提供一种更快、更通用的方法，可以显著改善新药的发现，以破坏通常与人类疾病相关的异常复合物。

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.