PROTEOMICS

蛋白质组学

• 批准号: 8378398
• 负责人: DIETER A WOLF
• 金额: $ 25.33万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-21 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8378398
• 关键词: Animal Model; Back; Biological; Biological Markers; Budgets; Cancer Center; Cancer Center Support Grant; Carbohydrates; Cell physiology; Cells; Complex; Computer software; Data Analyses; Databases; Descriptor; Diagnostic; Experimental Designs; Fractionation; Future; Gel; Genetic; Goals; High Pressure Liquid Chromatography; Human Resources; Institutes; Isotope Labeling; Label; Liquid Chromatography; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Medical Research; Messenger RNA; Methodology; Methods; Molecular Analysis; Molecular Profiling; Molecular Weight; Names; Neurosciences; Normal Cell; Pathway Analysis; Pathway interactions; Peptides; Phenotype; Phosphorylation; Post-Translational Protein Processing; Preparation; Protein Analysis; Proteins; Proteome; Proteomics; Proteomics Shared Resource; Qualifying; Research; Research Infrastructure; Research Personnel; Resolution; Resource Sharing; Sampling; Scientist; Series; Services; Solutions; Somatic Mutation; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; System; Technology; Testing; Tissues; United States National Institutes of Health; base; data management; design; experience; instrument; instrumentation; investigator training; malignant phenotype; mass spectrometer; meetings; member; new technology; new therapeutic target; novel; outreach; programs; protein complex; protein expression; protein function; protein profiling; protein protein interaction; tandem mass spectrometry

The Proteomics Shared Resource at Burnham Institute offers a state-of-the-art infrastructure for protein analysis at a global level that is tailored to the specific needs of Cancer Center investigators. The core technologies include methods for upfront sample fractionation and back-end protein identification by mass spectrometry (MS) and database searching. Facility MS instruments include Thermo LTQ and LTQ Orbitrap systems, and Bruker HCTUItra and MALDI Autoflex II TOF/TOF systems. Routine services include protein/peptide molecular weight determination by MALDI-MS, differential 2D-PAGE analysis of low to moderately complex samples, identification of gel separated proteins by tandem MS, and post-translational modification mapping for known peptides and proteins. More recently added services include analysis for complex protein samples by multidimensional liquid chromatography (MD LC-MS/MS), phosphoproteomics using 2D or 3D LC-MS/MS methodology, targeted quantitative proteomics employing isotope-labeling strategies (ICPL/ITRAQ/TM), and differential quantitative analysis of complex samples using label-free strategies. Cancer Center investigators are utilizing these technologies in a variety of ways, but most frequently to identify novel components of protein complexes and protein-protein interactions, to map functionally important post-translational modifications such as phosphorylation and ubiquitylation, and to comprehensively profile quantitative differences in protein abundance in cancer versus normal cells. To facilitate these projects, facility personnel routinely meet and interact with investigators to provide guidance with project design, samples preparation, and data analysis. In addition, the facility entertains a rich portfolio in outreach activities geared toward training investigators and their associates in proteomic technology. The facility's future activities will focus on maintaining high quality services as well as the exploration, testing, and demand-based implementation of novel technologies that will benefit the Center's research programs. Efforts in planning toward these goals include promoting additional technologies for accurate quantitative protein profiling in the low abundance range (SILAC), enhancing data analysis, data management and biological interpretation (pathway analysis), and expanding the facility's personnel and instrumentation capacity to a high resolution, high sensitivity mass spectrometry based proteomics facility. In the past year, 25 Cancer Center members used the Proteomics Shared Resource. The requested CCSG support in the first year of $168,553 is 16.6% of the total projected annual Proteomics Shared Resource budget.

伯纳姆研究所的蛋白质组学共享资源为蛋白质组学提供了最先进的基础设施。 在全球范围内进行分析，以满足癌症中心研究人员的特定需求。核心 技术包括前期样品分离和后期蛋白质质量鉴定方法 质谱（MS）和数据库搜索。设施MS仪器包括Thermo LTQ和LTQ Orbitrap 系统以及Bruker HCTUItra和MALDI Autoflex II TOF/TOF系统。常规服务包括 通过MALDI-MS测定蛋白质/肽分子量，低至 中度复杂的样品，通过串联MS鉴定凝胶分离的蛋白质，和翻译后 已知肽和蛋白质的修饰作图。最近增加的服务包括分析 多维液相色谱法（MD LC-MS/MS）分析复杂蛋白质样品，磷酸化蛋白质组学 使用2D或3D LC-MS/MS方法，采用同位素标记的靶向定量蛋白质组学 策略（ICPL/ITRAQ/TM），以及使用无标记的 战略布局癌症中心的研究人员正在以各种方式利用这些技术，但大多数 经常鉴定蛋白质复合物和蛋白质-蛋白质相互作用的新组分， 功能上重要的翻译后修饰，如磷酸化和泛素化，以及 全面地分析癌症与正常细胞中蛋白质丰度的定量差异。到 为了促进这些项目，设施人员定期与调查人员会面并进行互动，以提供指导 项目设计、样品制备和数据分析。此外，该设施还提供丰富的投资组合， 在面向培训研究人员和他们的同事在蛋白质组学技术的推广活动。的 设施的未来活动将集中在保持高质量的服务以及勘探，测试， 和基于需求的新技术的实施，这将有利于中心的研究计划。 为实现这些目标而进行的规划工作包括推广其他技术， 低丰度范围蛋白质谱分析（SILAC），加强数据分析、数据管理和 生物学解释（途径分析），并扩大设施的人员和仪器 高分辨率、高灵敏度质谱分析为基础的蛋白质组学设施的能力。在过去的一年里， 25名癌症中心成员使用了蛋白质组学共享资源。所要求的CCSG在 第一年的168，553美元是蛋白质组学共享资源预算总额的16.6%。