Ultraviolet Photodissociation Mass Spectrometry

Jennifer S.德克萨斯大学奥斯汀分校的Brodbelt得到化学系化学测量和成像项目的支持，开发基于质谱的分析方法，用于分析完整蛋白质。 这些实验提供了关于多个翻译后修饰的位置的信息，这使得有可能研究这些基团对蛋白质功能的影响，从而更好地理解生物系统的结构和性质。 拟议的研究有望克服与完整蛋白质的激活和解离相关的一些技术障碍，以生成可用于蛋白质鉴定的有意义的片段离子图谱。 如果成功的话，这项工作将对人类蛋白质组计划做出积极的贡献，这是一项正在进行的对所有人类蛋白质进行编目的工作。 它还将促进自上而下的蛋白质质谱法的使用，以解决生物治疗，结构生物学和机械生物化学领域的问题。 本研究课题的目的是开发紫外光解离（UVPD）技术，使其成为质谱分析蛋白质的通用方法，并探索在紫外光解离过程中发挥作用的机理、结构和实验特征。 该项目有五个独立但相关的具体目标：（1）探索完整蛋白质的有效UVPD参数;（2）开发电子转移解离-紫外光解离的混合方法（3）改变蛋白质的电荷状态以实现更有效的紫外光解离;（4）通过将特定发色团附着在蛋白质上来调节UVPD的选择性;（5）发展UVPD方法学在结构生物学中的应用，重点是确定完整蛋白质的结构和天然蛋白质复合物的结合相互作用。在本项目期间，学生将接受先进质谱，激光光学，衍生反应，蛋白质加工和生物信息学的培训，并将设立夏令靴子营，以提高对先进质谱在解决生物问题方面的潜在应用的认识。

Professor Jennifer S. Brodbelt of the University of Texas at Austin is supported by the Chemical Measurement and Imaging Program in the Division of Chemistry to develop analytical methodologies based on mass spectrometry for the analysis of intact proteins. These experiments provide information on the locations of multiple posttranslational modifications, which makes it possible to study the influence of those groups on protein function and hence allow better understanding of the structure and properties of biological systems. The proposed research promises to overcome some of the technical hurdles associated with the activation and dissociation of intact proteins to generate meaningful fragment ion maps that can be used for protein identification. If successful, the proposed work will positively contribute to the Human Proteome Project, an on-going effort to catalog all human proteins. It will also facilitate the use of top-down protein mass spectrometry for solving problems in the areas of biotherapeutics, structural biology, and mechanistic biochemistry. In this context, the project will provide new avenues for the identification of proteins and hence advance our understanding of life processes.In this project, studies are being conducted to develop ultraviolet photodissociation (UVPD) as a versatile strategy for the analysis of proteins by mass spectrometry, and to explore the mechanistic, structural, and experimental features that play a role in the ultraviolet photodissociation process. The project has five separate but related specific aims: (1) to explore the parameters for effective UVPD of intact proteins; (2) to develop a hybrid electron transfer dissociation- ultraviolet photodissociation (ETD/UVPD) methods for mass spectrometric fragmentation of intact proteins; (3) to modify the charge state of proteins for more effective ultraviolet photodissociation; (4) to modulate the selectivity of UVPD by the attachment of specific chromophores to the proteins; and (5) to develop UVPD methodologies for structural biology applications with a focus on determining the structures of intact proteins and the binding interactions of native protein complexes.During the course of this project, students will be trained in advanced mass spectrometry, laser optics, derivatization reactions, protein processing, and bioinformatics, and summer boot camps will be set up to increase awareness of the potential applications of advanced mass spectrometry for solving biological problems.