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.

本研究的目标是研究确定蛋白质结构、测量蛋白质的新方法蛋白质-蛋白质结构和结合，并利用质量分离和鉴定不同的蛋白质构象异构体光谱测定方法。将进行溶液相和气相研究。从差异来看这两个阶段的结构或结合相互作用，有关溶剂如何影响两种蛋白质的信息可以确定蛋白质之间的构象和特定的分子间相互作用。此信息可能会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测定方法。一种灵敏、高通量的蛋白质测定方法构象可以极大地提高研究人员发现蛋白质功能和识别新蛋白质的能力基于结构的药物。非共价配合物的串联质谱实验将调查了。这些研究可以提供其他方法难以或无法获得的结构信息方法。具体目标包括 1) 开发一种潜在灵敏且快速的蛋白质测定方法使用溶液相 H/D 交换和电子捕获解离来确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以及 3) 研究高场不对称波形离子淌度光谱学作为蛋白质构象分析的快速且灵敏的方法。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通过气相实验确定本体溶液中离子的结构。本研究旨在开发快速确定折叠结构的新方法蛋白质，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蛋白质互动。这些研究可以提供重要的新信息，有助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、海绵状病脑病（例如疯牛病或克雅氏病），甚至某些癌症。此外，蛋白质-蛋白质相互作用的研究可能提供一种更快、更通用的方法显着促进新药的发现，以破坏经常出现的异常复合物与人类疾病有关。