2D NANO-FLOW MASS SPECTROMETRY SYSTEM

该子项目是利用由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。子项目和研究者（PI）可能从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是用于资助的，不一定是研究者的机构。描述（由申请人提供）：加州圣地亚哥大学的生物分子质谱设施希望获得2D纳米流质谱仪系统。我们提出的配置包括LC Packings U-3000 Nano-LC（带开关）和Ultimate-3000双泵（用于2D色谱），与Finnigan LTQ（带Sequest Sorcerer数据分析集群）接口。这种组合具有所提出的研究所需的可靠性、高通量能力和灵敏度。该仪器将用于蛋白质组学研究，以了解特定基因产物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物，以确定翻译后修饰，并确定生物活性肽产生的途径。拟议研究项目的目标包括以稳健和全面的方式鉴定复合物中的蛋白质。特别是，我们将分析参与阿尔茨海默病，癌症和细菌发病机制的蛋白质复合物。此外，参与疼痛和血压控制的生物活性肽的生物合成途径的蛋白质组学，以及产生它们的蛋白酶将被进行。翻译后修饰的分析，开始与磷酸化的激酶锚定蛋白，也计划。最后，提出了一种新的方法来识别蛋白质，不一定直接结合在附近。教授E。Komives，J.狄克逊和S.泰勒将识别复合物中的蛋白质。胡克教授和D. T.奥康纳将分析生物活性肽的生物合成。R教授Tsien将探索使用ReAsH试剂发现细胞中紧邻蛋白质的可能性。

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.