Ultraviolet Photodissociation Mass Spectrometry

紫外光解离质谱法

基本信息

批准号：
1402753
负责人：
Jennifer Brodbelt
金额：
$ 49.5万
依托单位：
University of Texas at Austin
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：
2014-09-01 至 2022-05-31
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=1402753&HistoricalAwards=false
关键词：
Ultraviolet Photodissociation Mass Spectrometry

项目摘要

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.

德克萨斯大学奥斯汀分校的 Jennifer S. Brodbelt 教授在化学系化学测量和成像项目的支持下，开发基于质谱的分析方法，用于分析完整的蛋白质。这些实验提供了有关多个翻译后修饰位置的信息，这使得研究这些基团对蛋白质功能的影响成为可能，从而更好地了解生物系统的结构和特性。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识别的有意义的碎片离子图。如果成功，拟议的工作将为人类蛋白质组项目做出积极贡献，该项目是一项对所有人类蛋白质进行编目的持续努力。它还将促进使用自上而下的蛋白质质谱来解决生物治疗、结构生物学和机械生物化学领域的问题。在这种背景下，该项目将为蛋白质的鉴定提供新的途径，从而促进我们对生命过程的理解。在该项目中，正在进行研究以开发紫外光解离（UVPD）作为蛋白质质量分析的通用策略光谱测定法，并探索在紫外光解过程中发挥作用的机制、结构和实验特征。该项目有五个独立但相关的具体目标：（1）探索完整蛋白质有效 UVPD 的参数； (2) 开发用于完整蛋白质质谱裂解的混合电子转移解离-紫外光解离(ETD/UVPD)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到蛋白质上来调节 UVPD 的选择性； (5) 开发用于结构生物学应用的 UVPD 方法，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。在该项目期间，学生将接受高级质谱、激光光学、将设立衍生反应、蛋白质加工和生物信息学以及夏季训练营，以提高人们对先进质谱技术在解决生物问题方面的潜在应用的认识。