2D NANO-FLOW MASS SPECTROMETRY SYSTEM

这个子项目是利用由NIH/NCRR资助的共享仪器赠款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是用于拨款的，而不一定是用于调查人员的机构。描述(由申请人提供)：加州大学圣地亚哥分校的生物分子质谱学设施希望获得2D纳米流动质谱计系统。我们建议的配置包括带有开关的U-3000 Nano-LC和用于2D层析的Ulal-3000双泵与带有SEQUEST Magcerer数据分析集群的Finnigan LTQ接口。这种组合具有拟议研究所需的可靠性、高通量能力和灵敏度。该仪器将用于蛋白质组学研究，以了解特定基因产品如何影响细胞功能和人类健康。质谱仪系统将用于发现相互作用复合体，识别翻译后修饰，并识别生物活性多肽的产生途径。拟议研究项目的目标包括以一种强有力和全面的方式识别复合体中的蛋白质。特别是，我们将分析阿尔茨海默病、癌症和细菌发病机制中涉及的蛋白质复合体。此外，还将进行与疼痛和血压控制有关的生物活性多肽的生物合成途径的蛋白质组学研究，以及产生它们的蛋白酶的蛋白质组学研究。对翻译后修饰的分析也在计划中，首先是激酶锚定蛋白的磷酸化。最后，提出了一种通过不一定直接结合来识别接近的蛋白质的新方法。E.Komives、J.Dixon和S.Taylor教授将识别复合体中的蛋白质。V.Hook和D.T.O‘Connor教授将分析生物活性多肽的生物合成。钱存训教授将探索使用ReAsH试剂发现细胞中近距离蛋白质的可能性。

This subproject is one of many research subprojects utilizing the resources provided by a Shared Instrumentation Grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the grant, which is not necessarily the institution for the investigator. DESCRIPTION (provided by applicant): The Biomolecular Mass Spectrometry Facility at the University of California San Diego wishes to acquire a 2D nano-flow mass spectrometer system. The configuration we propose consists of an LC Packings U-3000 Nano-LC with Switches and Ultimate-3000 Dual pumps for 2D chromatography interfaced to a Finnigan LTQ with Sequest Sorcerer data analysis cluster. This combination has the reliability, high-throughput capabilities, and sensitivity required for the proposed studies. This instrument will be used in proteomics studies to understand how specific gene products work to impact cellular function and human health. The mass spectrometer system will be used to discover interaction complexes, to identify post-translational modifications, and to identify pathways of bioactive peptide generation. The goals of the proposed research projects include identification of proteins in complexes in a robust and comprehensive way. In particular, we will analyze protein complexes involved in Alzheimer's Disease, cancer, and bacterial pathogenesis. Also, proteomics of the biosynthetic pathways of bioactive peptides involved in pain and blood pressure control, and the proteases that generate them will be carried out. Analysis of post-translational modifications, beginning with phosphorylation of the kinase anchoring proteins, is also planned. Finally, a novel approach to identifying proteins that are in close proximity by not necessarily binding directly is proposed. Professors E. Komives, J. Dixon and S. Taylor will identify proteins in complexes. Professors V. Hook and D. T. O'Connor will analyze the biosynthesis of bioactive peptides. Professor R. Tsien will explore the possibilities of using ReAsH reagents to discover proteins in close proximity in the cell.