Protein Conformation and Noncovalent Interactions

描述（由申请人提供）：探测结构的准确方法， 生物聚合物的构象和动力学，以及它们特定的非共价键 相互作用，对于理解生物化学是非常重要的， 分子水平。质谱法具有快速、准确、可靠等优点， 灵敏度（阿托摩尔检测）和高特异性（质量准确度+/-几个 Da> 100，000Da）。串联质谱法用于鉴定和 定位翻译后修饰和结合位点， 存在于复杂的混合物中，等等。 使用电子捕获的主链断裂的效率和程度 解离，并发展这种方法来获得有关生物聚合物的信息 构象在气相和溶液相中的H/D交换测量， 将结合碰撞截面测量，电子捕获 解离和分子动力学模拟，以获得详细的信息 关于溶液和气相中的构象。通过比较结果 在这两种不同的环境中，关于溶剂对 可以直接获得生物聚合物构象。另一个目的是了解 离子结构在从本体溶液过渡到 电喷雾电离过程中气相的隔离环境 过程希望通过了解这一过程，我们可以将 生物聚合物和非共价复合物的结构信息， 气相实验回到本体溶液中离子的结构。 另一个目的是了解特定的非共价相互作用的作用， 溶剂对这些相互作用的影响。非共价相互作用是 在酶-底物反应性、超分子复合物形成和 在许多疾病中，例如基于朊病毒的疾病， 脑病和克罗伊茨雅氏病。

DESCRIPTION (provided by applicant): Accurate methods to probe the structure, conformation and dynamics of biopolymers, as well as their specific noncovalent interactions, are tremendously important for understanding biochemistry at the molecular level. Mass spectrometry has the advantages of high speed, ultrahigh sensitivity (attomole detection), and high specificity (mass accuracy +/- a few Da at >100,000 Da). Tandem mass spectrometry is used for identifying and locating postranslational modifications and binding sites, identifying proteins present in complex mixtures, etc. It is one aim of this proposal to improve the efficiency and extent of backbone fragmentation using electron capture dissociation, and develop this method to obtain information about biopolymer conformation. H/D exchange measurements, both in the gas and solution phase, will be combined with collisional cross section measurements, electron capture dissociation, and molecular dynamics simulations to obtain detailed information about conformation both in solution and in the gas phase. By comparing results in these two different environments, information about the role of solvent on biopolymer conformation can be directly obtained. Another aim is to understand how the structure of an ion changes during the transition from bulk solution to the isolated environment of the gas phase during the electrospray ionization process. It is hoped that by understanding this process, we can relate the structural information of biopolymers and noncovalent complexes determined from gas-phase experiments back to the structures of the ions in bulk solution. Another aim is to understand the role of specific noncovalent interactions and the influence of solvent on these interactions. Noncovalent interactions are important in enzyme-substrate reactivity, supermolecular complex formation and in many diseases, such as prion based diseases including spongiform encephalopathies and CreutzfeldJacob disease.