Development of New Approaches and Methods in Biological Mass Spectrometry and Proteomics.

生物质谱和蛋白质组学新途径和新方法的发展 **我们的研究计划涉及生物质谱和蛋白质组学的新方法和新方法的开发，以及它们在解决生物学问题中的应用。 我们的研究是高度多学科的，涉及质谱，生物信息学，分离科学以及生物和生物医学科学的各个领域。** 尽管质谱和相关技术取得了重大技术进步，人类蛋白质组的初稿也已出版，但仍有许多尚未解决的问题需要解决，以解决复杂的生物学问题。剩下的挑战包括低丰度蛋白质的检测和翻译后修饰的全面表征和定量。** 在本资助申请中，我们提出了一些创新策略，通过使用我们实验室中最先进的技术和仪器（LTQ Orbitrap Elite，Q-Exactive，4000 Q-Trap）来解决这些具体问题。 这项研究的一个重要组成部分将涉及制备合成肽作为标准品和开发生物信息学工具，以促进复杂质谱数据的分析。(e.g.,基于精确质量和LC保留时间的蛋白质鉴定;根据含有一种以上肽的MS/MS光谱进行鉴定）。为了确定蛋白质翻译后修饰的完整程度，我们将使用“自上而下”和“中等自下”的方法与传统的“自下而上”的蛋白质组学相结合。为了做到这一点，我们将使用新的酶和新的条件，以达到最佳的大小 * 的分析。** 这项研究将使学生接触到各种技术，并将为他们的培训提供肥沃的环境。

Development of New Approaches and Methods in Biological Mass Spectrometry and Proteomics.******Our research program pertains to the development of new approaches and new methods in biological mass spectrometry and proteomics as well as their applications to solve biological questions. Our research is highly multidisciplinary involving the fields of mass spectrometry, bioinformatics, separation sciences and various areas in the biological and biomedical sciences. ******Despite major technical advances in mass spectrometry and associated technologies and the publication of first drafts of the human proteome, there are still many unresolved issues that needed to be addressed to solve complex biological questions. Remaining challenges include the detection of low abundance proteins and the full characterization and quantification of post-translational modifications. *******In this grant application we propose a number of innovative strategies to address these specific issues by using state-of-the-art techniques and instrumentation available in our laboratory (LTQ Orbitrap Elite, Q-Exactive , 4000 Q-Trap). An important component of this research will involve preparation of synthetic peptides as standards and the development of bioinformatics tools to facilitate the analysis of complex mass spec data. (e.g., protein identification based on accurate mass and LC retention time; identification from MS/MS spectra containing more than one peptide). In order to identify the full extent of post-translational modifications of proteins we will use "top-down" and "medium-down" approaches in combination with traditional "bottom-up" proteomics. To do this we will use new enzymes and new conditions to achieve optimal size***for analysis. ******This research will expose students to a variety of techniques and will be a fertile environment for their training.