La Jolla Interdisciplinary Neuroscience Center Cores - Pro Core BU

蛋白质组学核心 一、概况和主要目标 神经科学蛋白质组学设施的任务是提供鉴定、表征、 并详细分析蛋白质、多肽、碳水化合物和类药物小分子。这个设施 将允许神经科学家访问La Jolla Torrey，从而促进他们之间的合作 Pines Mesa(主要由伯纳姆、索尔克、斯克里普斯和加州大学圣迭戈分校的神经科学家组成)。该设施 将利用伯纳姆医学研究所的现有基础设施和知识库 Research(BIMR)现有的蛋白质组学中心以及最先进的质谱学(MS)，请访问 斯克里普斯研究所(TSRI)和索尔克研究所，它们组成了位于 街对面是拉霍亚Torrey Pines北路上的BIMR(距离加州大学圣地亚哥分校不到半英里)。 目前，BIMR的这个核心设施是伯纳姆国立卫生研究院资助的癌症中心的一部分，几乎没有 访问La Jolla Torrey Pines台地的神经科学家。根据目前的NIH申请 为神经科学中心核心提供蓝图赠款，所有现代蛋白质组技术将可用于 该设备的神经科学用户包括复杂蛋白质组的全面蛋白质组学分析 使用多维蛋白质鉴定(MUDIT)、基于活性的酶化学图谱 用于识别蛋白酶(产品)底物的质量标签、产品终端同位素编码 蛋白质分解)，利用前体离子扫描鉴定磷酸肽和蛋白质，中性损失 扫描和MS3扫描，以及自动高通量MALDI-TOF/TOF分析蛋白质和 多肽混合物。该设施还对完整的重组人进行更常规的质量分析 蛋白质、合成肽和糖肽，从凝胶切片/斑点中鉴定蛋白质 多肽质量指纹图谱和多肽测序。2006年春，该基金主任将 也申请资金购买将提供最高质量精度和分辨率的FTMS， 广泛的分段能力(如CID、ECD和IRMPD)和从头执行的能力 多肽和蛋白质测序(自上而下的蛋白质组学)，并直接表征最复杂的 完整蛋白质的翻译后修饰，无需样品的事先消化和处理。 该奖项的一个主要目标是整合La Jolla神经生物学的蛋白质组学工作 伯纳姆医学研究所现有的蛋白质组学中心。这将是 涉及购买额外的仪器(在第一年购买LTQ离子陷阱)，以及雇用 额外的人员。通过融合神经科学蛋白质组学共享的资源 当前中心(包括伯纳姆癌症中心的蛋白质组学设施以及 NTCNP蛋白质分解途径中心)，我们将能够利用相当大的 现有中心的仪器和专业知识，以造福神经科学界。这个 目前的拨款申请将促进神经科学家平等使用这些设施，而不是 目前可以进入，因为设施在癌症中心。神经科学的整合 进入现有蛋白质组学中心的蛋白质组学工作将为 神经科学蛋白质组学研究。除了伯纳姆研究所现有的设备外， 上面列出的医学研究(BIMR)，是在Jeff Smith博士的指导下，卫星设施 在索尔克研究所(在沃尔夫冈·费舍尔的指导下)和斯克里普斯也有 在著名蛋白质组学权威机构约翰·耶茨的指导下，TSRI，III。

PROTEOMICS CORE 1. OVERVIEW AND MAIN OBJECTIVES The mission of the Neuroscience Proteomics Facility is to provide identification, characterization, and detailed analysis of proteins, peptides, carbohydrates and drug-like small molecules. This facility will allow access to and thereby foster collaborations among Neuroscientists on the La Jolla Torrey Pines Mesa (comprised primarily of Burnham, Salk, Scripps, and UCSD neuroscientists). The facility will leverage the existing infrastructure and knowledge-base of the Burnham Institute for Medical Research's (BIMR's) existing Proteomics Center as well as state-of-the-art mass spectrometry (MS) at The Scripps Research Institute (TSRI) and The Salk Institute, which constitute Satellite Centers located across the street from BIMR on North Torrey Pines Road in La Jolla (and within half a mile from UCSD). Presently this Core Facility at BIMR is part of the Burnham's NIH-funded Cancer Center with little or no access to neuroscientists on the La Jolla Torrey Pines Mesa. Under the present application for an NIH Blueprint Grant for Neuroscience Center Cores, all modern proteomic technologies will be available to neuroscience users of the Facility including comprehensive proteomic analysis of complex proteomes using Multi-dimensional Protein Identification (MUDPIT), activity-based enzyme profiling with chemical mass tags, Product Terminal Isotope Coding for the identification of substrates for proteases (products of proteolysis), phospho-peptide and protein identification using precursor ion scanning, neutral loss scanning and MS3 scanning, and automated high-throughput MALDI-TOF/TOF analysis of protein and peptide mixtures. The Facility also performs the more routine mass analysis of intact recombinant proteins and synthetic peptides and glycopeptides, protein identification from gel slices/spots using peptide mass fingerprinting and peptide sequencing. In the spring of 2006 the Director of the Facility will also apply for funds to purchase an FTMS which will provide the highest mass accuracy and resolution, broad fragmentation capability (such as CID, ECD and IRMPD) and the ability to perform de novo peptide and protein sequencing (top-down proteomics) and directly characterize the most complex post-translational modifications in intact proteins, without prior digestion and processing of the sample. A primary objective for this award is to integrate the proteomics efforts of the La Jolla Neurobiology community into the existing Proteomics Center at the Burnham Institute for Medical Research. This will involve the purchase of additional instrumentation (an LTQ Ion Trap in Year 1), and the hiring of additional personnel. By amalgamating the neuroscience Proteomics Shared Resource with that of the current Center (which includes the Proteomics Facility for the Burnham's Cancer Center as well as the NTCNP Center on Proteolytic Pathways), we will be able to capitalize on the considerable instrumentation and expertise of the existing Center for the benefit of the Neuroscience community. The current grant application will facilitate equal access to these facilities by Neuroscientists, who do not have access currently because the facility is in the Cancer Center. The integration of the Neuroscience proteomics effort into the existing Proteomics Center will provide a substantial jump-start for the Neuroscience proteomics effort. In addition to the current equipment at the Burnham Institute for Medical Research (BIMR) listed above, which is under the direction of Dr. Jeff Smith, satellite facilities are also available at The Salk Institute (under the direction of Wolfgang Fischer) and at The Scripps Research Institute (TSRI), under the direction of noted proteomics authority John Yates, III.