Transient Intermediates in Proton and Electron Transfer

在这个由化学部化学结构、动力学机制B项目资助的项目中，西雅图华盛顿大学化学系的Frantisek Turecek教授将开发通过质谱法改进蛋白质测序的新方法。 本研究的目标是首先深入了解肽骨架解离，提供肽氨基酸序列的信息，然后拼接以完成亲本蛋白质的序列。蛋白质测序是确定导致许多疾病和退行性过程的突变、翻译后修饰和损伤的关键步骤。癌症、帕金森和阿尔茨海默病以及衰老都包括蛋白质变化，这是破坏细胞过程及其调节的主要原因之一。该研究小组也有能力为不同文化背景的学生提供最高水平的教育和培训，包括那些在物理和生命科学领域代表性不足的学生。 质谱法是蛋白质和肽分析的首选方法。在所谓的自下而上方法中，蛋白质或其混合物被酶促切割成肽的混合物，所述肽的混合物被在线分离并分析。质谱分析提供了关于肽质量和氨基酸序列和组成的有价值的信息。可以通过与数据库匹配来评估信息，以找到存在已知基因的通用蛋白质。然而，来自缺乏基因组数据的生物体的蛋白质或由于疾病或损伤而修饰的蛋白质必须通过所谓的从头测序来分析，这意味着仅从质谱数据读取完整的序列。 在这个项目中，研究和开发新的方法来改善从头蛋白质测序。 该项目有几个目标：（1）正在开发一种在固相支持物上进行肽捕获和化学修饰的新方法，以简化肽混合物并提高序列读出水平。(2)将进行电子转移后肽离子解离的基础研究，以确定离子3D结构和能量沉积的影响。(3)介绍了一种新的紫外光解离方法，以提供一种不同的活化手段，引起气相肽离子的有效解离。这些研究的结果将用于完善目前的肽离子活化和解离的理论模型，并利用它来预测离子行为。

In this project funded by the Chemical Structure, Dynamic & Mechanism B Program of the Chemistry Division, Professor Frantisek Turecek of the Department of Chemistry at University of Washington, Seattle, will develop new methods for improved protein sequencing by mass spectrometry. The goal of this research is first to gain deep understanding of peptide backbone dissociations that provide information on the peptide amino acid sequence which is then spliced to complete the sequence of the parent protein. Protein sequencing is the critical step in determining mutations, post-translational modifications, and damage that cause many diseases and degenerative processes. Cancer, Parkinson and Alzheimer diseases and aging all include protein changes as one of the major causes disrupting cellular processes and their regulation. This research group is also well-positioned to provide the highest level of education and training for students of different cultural backgrounds including those underrepresented in physical and life sciences. Mass spectrometry is the method of choice for protein and peptide analysis. In the so-called bottom-up approach, the protein or a mixture thereof is enzymatically cleaved to a mixture of peptides that are separated on-line and analyzed. Mass spectrometry analysis provides valuable information on the peptide mass and amino acid sequence and composition. The information can be evaluated by matching against a data base to find the generic protein for which there is a known gene. However, proteins from organisms lacking genome data, or modified proteins due to disease or damage must be analyzed by the so-called de novo sequencing, implying complete sequence readout from the mass spectrometry data alone. In this project, new methods are investigated and developed to improve de novo protein sequencing. The project has several goals: (1) A new method of peptide capture and chemical modification on solid phase support is being developed to simplify peptide mixtures and increase the level of sequence readout. (2) Fundamental studies of peptide ion dissociations following electron transfer will be carried out to determine the effects of ion 3D structure and energy deposition. (3) A new UV photodissociation method is introduced to provide a different means of activation, causing efficient dissociation of gas-phase peptide ions. Results from these studies will be used to refine the current theoretical model of peptide ion activation and dissociation and utilize it for prediction of ion behavior.