Biomolecular Characterization via Radical Directed Dissociation

教授瑞安R.加州大学滨江分校的Julian得到了生物科学理事会化学部化学测量和成像（CMI）计划和生物研究仪器开发（IDBR）计划的支持，以进一步开发他的质谱方法，这些方法最初是为肽和蛋白质的鉴定和表征而开发的。该项目将扩大质谱法可应用于生物分子的范围，为分子生物学、生物化学和医学等众多领域的科学研究开辟新途径。 具体而言，所提出的用于改进寡糖分析的方法注定是广泛有用的，因为这些是生物学和生物医学上重要的分子，现有工具对其仅适度有用。 在这方面，该项目有望提高我们对生命过程的理解。在进行这项研究的过程中，研究生和本科生将获得生物分析技术，质谱和化学合成方面的宝贵技能。 PI将积极参与UCR计划（GradEDge和AGEP），专注于研究生院内少数民族学生的成功发展，研究团队将参加当地K-12学校的科学展览和科学演示该项目有三个具体目标：（1）探索自由基定向解离（Radical Directed Dissociation，RDD）技术在寡糖结构解析中的应用，并阐明RDD在寡糖中产生的独特裂解途径。 该目标的重点将是一系列靶点，包括糖胺聚糖、O-和N-连接聚糖和糖肽;（2）使用RDD鉴定抗氧化肽并测定其溶液相抗氧化能力。 生物学相关蛋白质的固有抗氧化能力和影响抗氧化能力的因素将被调查，序列取代和MS/MS实验的组合将被用来揭示有关自由基是如何被隔离的信息;（3）利用光笼共价标记和光化学映射蛋白质，特别是多蛋白复合物的高阶溶液相结构。

Professor Ryan R. Julian of the University of California Riverside is supported by the Chemical Measurement and Imaging (CMI) Program in the Division of Chemistry and the Instrument Development for Biological Research (IDBR) Program in the Directorate for Biological Sciences to further develop his mass spectrometry methodologies, which were initially developed for the identification and characterization of peptides and proteins. The project will expand the scope of biomolecules to which mass spectrometry can be applied, opening up new avenues of scientific research in numerous fields including molecular biology, biochemistry and medicine. Specifically, the proposed methods for improved oligosaccharide analysis are destined to be broadly useful as these are biologically and biomedically important molecules for which existing tools are only moderately useful. In this context, the project promises to enhance our understanding of life processes. In the course of conducting this research, graduate and undergraduate students will acquire valuable skills in bioanalytical technologies, mass spectrometry and chemical synthesis. The PI will actively participate in UCR programs (GradEDge and AGEP) that are focused on the successful advancement of minority students within the graduate school, and the research team will participate in science fairs and science demonstrations at local K-12 schoolsThe project is organized in three specific objectives: (1) to explore the utility of radical directed dissociation (RDD) for the elucidation of the structure of oligosaccharides and to delineate the unique fragmentation pathways that RDD produce in oligosaccharides. The focus of this objective will be on a range of targets including glycosaminoglycans, O- and N- linked glycans and glycopeptides; (2) to use RDD to identify antioxidant peptides and determine their solution phase antioxidant capacity. The inherent antioxidant capacity of biologically relevant proteins and the factors that influence antioxidant capacity will be investigated, and a combination of sequence substitution and MS/MS experiments will be used to reveal information about how radicals are sequestered; and (3) to utilize photocaged covalent labels and photochemistry to map the higher order solution phase structure of proteins, in particular multiprotein complexes.