Ultraviolet Photodissociation Mass Spectrometry

在化学系化学测量和成像项目的支持下，德克萨斯大学奥斯汀分校的Jenny Brodbelt教授和她的团队正在开发利用质谱（MS）表征蛋白质的创新方法。细胞中的蛋白质参与维持生命所必需的大量功能，蛋白质的生理功能和活动是通过结构修饰来调节的。Brodbelt博士和她的团队开发了新的高性能方法，将质谱和气相分离和反应相结合，以检查蛋白质并确定分子细节上的修饰。特别是，Brodbelt小组使用高能激光粉碎蛋白质，以创建诊断碎片模式，揭示结构和修饰。该综合计划让学生在多个层面上参与合作、跨学科的研究，并为他们向科学事业的过渡提供专业培训。完整蛋白质的分析仍然是蛋白质组学的前沿之一，而描述完整蛋白质的能力为绘制在蛋白质结构和功能中起关键作用的组合修饰提供了最好的手段之一。在过去的十年中，broadbelt小组的一个中心标志是紫外光解（UVPD）在生物分子分析中的发展和应用。本项目重点介绍了UVPD在完整蛋白分析中的发展和应用。主要目的是整合动态离子操作策略，利用质子转移反应，气相分馏和电荷检测测量方法来扩大完整蛋白质的MS/MS光谱的信息含量。UVPD被用来表征异构体的离子迁移率通过模块漂移细胞分离，并测量碰撞截面的蛋白质。这些创新的方法被应用于破译蛋白质翻译后修饰的模式，并扩大了自上而下的蛋白质组学的质量范围。培训和指导活动与研究生和本科生建立伙伴关系，鼓励他们在学术和工业领域的专业发展成为未来的领导者。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

With support from the Chemical Measurement and Imaging program in the Division of Chemistry, Professor Jenny Brodbelt and her group at the University of Texas at Austin are developing innovative methods to characterize proteins using mass spectrometry (MS). Proteins in cells participate in a vast array of functions necessary to sustain life, and the physiological functions and activities of proteins are modulated by structural modifications. Dr. Brodbelt and her group develop new high performance methods that combine MS and both gas-phase separations and reactions to examine proteins and pinpoint the modifications in molecular detail. In particular, the Brodbelt group uses high energy lasers to shatter proteins to create diagnostic fragmentation patterns that reveal structures and modifications. The comprehensive plan engages students at multiple levels in collaborative, interdisciplinary research and provides professional training for the transition to scientific careers. Analysis of intact proteins remains one of the frontiers of proteomics, and the ability to characterize intact proteins offers one of the best means to map combinatorial modifications that play critical roles in protein structure and function. A central hallmark of the Brodbelt group over the past decade is the development and application of ultraviolet photodissociation (UVPD) for analysis of biological molecules. This project emphasizes the development and applications of UVPD for analysis of intact proteins. The main aims integrate dynamic ion manipulation strategies utilizing proton transfer reactions, gas-phase fractionation, and charge detection measurement methods to amplify the information content of MS/MS spectra of intact proteins. UVPD is used to characterize isomers separated by ion mobility via a modular drift cell, and collision cross-sections of proteins are measured. These innovative methods are applied to decipher patterns of post-translational modifications of proteins and extend the mass range of top-down proteomics. The training and mentoring activities forge partnerships with graduate and undergraduate students to encourage their professional development into future leaders in academic and industrial sectors.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.