Protein Conformation and Noncovalent Interactions

蛋白质构象和非共价相互作用

基本信息

批准号：
7577394
负责人：
Evan R Williams
金额：
$ 26.98万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-02-01 至 2011-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7577394
关键词：
Active Sites Affect Affinity Alzheimer&apos s Disease Amino Acids Apoptosis Back Binding Binding Proteins Biology Biopolymers Cattle Chemicals Clinical Complex Computing Methodologies Creutzfeldt-Jakob Syndrome Crystallography Cystic Fibrosis Disease Dissociation Electrons Elements Gases Goals Individual Intervention Ions Libraries Malignant Neoplasms Maps Mass Spectrum Analysis Measurement Measures Medicine Methods Modeling Molecular Conformation Pharmaceutical Preparations Phase Play Prion Diseases Process Protein Binding Protein Conformation Proteins Proteomics Reagent Research Research Personnel Resolution Role Sampling Signal Transduction Site Solutions Solvents Spectrum Analysis Structure Tube Water Work base conformer drug discovery enzyme activity high throughput analysis human disease improved inhibitor/antagonist intermolecular interaction ion mobility molecular assembly/self assembly molecular size mutant physical property protein complex protein function protein misfolding protein protein interaction protein structure rapid technique receptor binding research study small molecule tandem mass spectrometry

项目摘要

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.

这项研究的目标是研究确定蛋白质结构，测量的新方法蛋白质蛋白质结构和结合，并使用质量分离并识别不同的蛋白质构象体光谱法。将进行溶液相和气相研究。来自差异在这两个阶段中的结构或结合相互作用，有关溶剂如何影响蛋白质的信息可以确定蛋白质之间的构象和特定分子间相互作用。此信息有可能增强用于确定蛋白质结构和折叠以及质量的计算方法药物发现的光谱法。确定蛋白质的敏感，高的方法构象可以极大地提高研究人员发现蛋白质功能并识别新的功能的能力基于结构的药物。非共价配合物的串联质谱实验将是调查。这些研究可以提供其他难以或无法获得的结构信息方法。具体目的包括1）开发一种潜在敏感和快速的方法来确定蛋白质使用溶液相H/D交换与电子捕获解离以识别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作为蛋白质构象分析的快速而敏感的方法。希望这些研究将为联系生物聚合物和非共价复合物的结构信息提供牢固的基础从气相实验中确定回到大体溶液中离子的结构。这项研究旨在开发新方法，以快速确定蛋白质，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互动。这些研究可以提供重要的新信息，这对于理解很有用其中蛋白质折叠率不良的疾病，包括阿尔茨海默氏病，囊性纤维化，海绵状脑病（例如，疯牛或克鲁兹菲尔特雅各布疾病），甚至是一些癌症。另外，蛋白质蛋白质相互作用的研究可能会提供更快，更通用的方法显着改善发现新药的发现，以破坏经常是的异常复合物与人类疾病有关。