Transient Intermediates in Proton and Electron Transfer

质子和电子转移中的瞬态中间体

• 批准号: 1359810
• 负责人: Frantisek Turecek
• 金额: $ 48万
• 依托单位: University of Washington
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-15 至 2018-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1359810&HistoricalAwards=false
• 关键词: Transient; Intermediates; Proton; Electron; Transfer

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.

在这个由化学部化学结构、动态和机制B项目资助的项目中，西雅图华盛顿大学化学系的Frantisek Turecek教授将开发新的方法来改进蛋白质的质谱测序。本研究的目的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的序列。蛋白质测序是确定突变、翻译后修饰和导致许多疾病和退行性过程的损伤的关键步骤。癌症、帕金森病、阿尔茨海默病和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。这个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。质谱法是蛋白质和多肽分析的首选方法。在所谓的自下而上的方法中，蛋白质或其混合物被酶裂解成在线分离和分析的肽混合物。质谱分析提供了有关肽质量和氨基酸序列和组成的有价值的信息。这些信息可以通过与数据库进行匹配来评估，以找到已知基因的一般蛋白质。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进行分析，这意味着仅从质谱数据就可以读取完整的序列。在这个项目中，研究和开发了新的方法来提高从头蛋白测序。该项目有几个目标：(1)正在开发一种新的肽捕获和固相载体化学修饰方法，以简化肽混合物并提高序列读出水平。(2)开展电子转移后肽离子解离的基础研究，确定离子三维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，提供了一种不同的激活手段，使气相肽离子有效解离。这些研究的结果将用于完善当前肽离子活化和解离的理论模型，并利用它来预测离子的行为。