PROTEOMICS

伯纳姆研究所的蛋白质组学共享资源为蛋白质提供最先进的基础设施 针对癌症中心研究人员的特定需求，在全球范围内进行分析。其核心是 技术包括前端样品分离和后端蛋白质质谱仪鉴定方法 光谱(MS)和数据库搜索。设施MS仪器包括Thermo LTQ和LTQ Orbitrap 以及Bruker HCTUItra和MALDI Autoflex II TOF/TOF系统。日常服务包括 MALDI-MS测定蛋白质/多肽分子量，低分子量差分2D-PAGE分析 中等复杂样品、凝胶分离蛋白质的串联质谱仪鉴定和翻译后分析 已知肽和蛋白质的修饰图谱。最近增加的服务包括分析 多维液相色谱(MD LC-MS/MS)、磷蛋白质组学研究 使用2D或3D LC-MS/MS方法，使用同位素标记的靶向定量蛋白质组学 策略(ICPL/iTRAQ/TM)和使用无标记的复杂样品的差示定量分析 战略。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 经常识别蛋白质复合体和蛋白质相互作用的新成分，以绘制地图 功能上重要的翻译后修饰，如磷酸化和泛素化，以及 全面剖析癌症细胞与正常细胞蛋白质丰度的定量差异。至 为这些项目提供便利，设施人员定期与调查人员会面并互动，以提供指导 具有项目设计、样品准备和数据分析的能力。此外，该设施还提供丰富的产品组合 在旨在对研究人员及其同事进行蛋白质组技术培训的外联活动中。这个 设施未来的活动将专注于保持高质量的服务以及勘探、测试、 以及基于需求的新技术的实施，这将使该中心的研究计划受益。 为实现这些目标而进行规划的努力包括促进更多的技术，以实现准确的定量 低丰度范围内的蛋白质图谱(SILAC)，增强数据分析、数据管理和 生物解释(路径分析)以及扩大设施的人员和仪器 拥有基于高分辨率、高灵敏度的蛋白质组学设备的能力。在过去一年里， 25名癌症中心成员使用蛋白质组学共享资源。中请求的CCSG支持 第一年为168,553美元，占蛋白质组学年度共享资源预算总额的16.6%。

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.