PNNL Proteome Characterization Center

项目摘要 PNNL蛋白质组表征中心（PCC）的总体目标是整合基因组信息 利用蛋白质组学技术全面表征蛋白质组和翻译后修饰 （PTM）以及癌症信号网络中蛋白质的功能相互作用。作为一个PCCs， 目前的临床蛋白质组肿瘤分析联盟（CPTAC）计划，PNNL成功地应用于 先进的蛋白质组学能力和专业知识，以全面的蛋白质组学和磷酸化蛋白质组学 卵巢高级别浆液性癌的特征来自癌症基因组图谱， 新的质谱方法，以促进分析完整的蛋白质，低化学计量PTM， 非常小的样本。计划中的民族解放国家警察委员会将在这些成就的基础上再接再厉， 通过表征CPTAC分配的多种新肿瘤类型， 经过验证的标准化、多重发现和有针对性的确证工作流程，以及引入 新的技术和平台，以改善蛋白质组学测量。人力 将分析NCI提供的生物标本和临床前模型。 发现研究部门将全面表征蛋白质组和多个PTM-组， 采用经验证的先进蛋白质组学技术， 平台，集成的工作流程和信息学工具，使用高度复用的同量异位素标记和状态， 最先进的液相色谱-串联质谱仪器，最多可处理300个样品 每一年。验证性研究组将验证基因组变异对 蛋白质组和PTM-组与经验证的靶向蛋白质组学平台（例如，多反应监测）， 肽标准和质量保证。多达100种高特异性、多重靶向蛋白质组学检测， 每年都将开发蛋白质和PTM，用于使用多达150个样本的确证性研究， 年我们还将在PNNL PCC完善和验证几个发展战略和平台 利用我们在超快气相分离和小体积等领域的持续技术开发， 规模的样品分析平台和方法，以进一步提高在这两个通量和灵敏度 发现和验证性研究。 PNNL PCC将与其他PCC，蛋白基因组数据分析中心和 CPTAC数据整合和生物信息学网络中的蛋白质基因组转化研究中心 分析，以及翻译应用。

PROJECT SUMMARY The overall objective of the PNNL Proteome Characterization Center (PCC) is to integrate genomic information with proteomic technologies to comprehensively characterize the proteome and post-translational modifications (PTMs) as well as the functional interaction of proteins in signaling networks in cancers. As one of the PCCs in the current Clinical Proteomic Tumor Analysis Consortium (CPTAC) program, PNNL successfully applied advanced proteomics capabilities and expertise to the comprehensive proteogenomic and phosphoproteomic characterization of ovarian high-grade serous carcinomas from The Cancer Genome Atlas, and has developed novel mass spectrometric approaches to facilitate the analysis of intact proteins, low stoichiometry PTMs, and extremely small samples. The planned PNNL PCC will build on these achievements and contribute to the success of the CPTAC network by characterizing multiple new tumor types assigned by CPTAC, using both the proven standardized, multiplexed discovery and targeted confirmatory workflows as well as introducing novel technologies and platforms, as appropriate, to improve proteomic measurements. Both human biospecimens and preclinical models provided by NCI will be analyzed. The Discovery Research Arm will comprehensively characterize the proteome and multiple PTM-omes in human biospecimens and preclinical samples of 2-3 cancers with the validated advanced proteomics platforms, integrated workflows and informatics tools, using highly multiplexed isobaric labeling and state-of- the-art liquid chromatography-tandem mass spectrometry instrumentation at a throughput of up to 300 samples per year. The Confirmatory Research Arm will verify the impact of genomic variation on the changes in proteome and PTM-omes with validated targeted proteomics platforms (e.g., multiple reaction monitoring), peptide standards and quality assurance. Up to 100 highly specific, multiplexed targeted proteomics assays for both proteins and PTMs will be developed each year for the confirmatory studies using up to 150 samples per year. We will also refine and validate several developmental strategies and platforms at the PNNL PCC leveraging our on-going technology development in areas such as ultrafast gas phase separations and small- sized sample analysis platforms and methodologies to further improve the throughput and sensitivity in both the discovery and confirmatory studies. The PNNL PCC will work closely with the other PCCs, Proteogenomic Data Analysis Centers and Proteogenomic Translational Research Centers in the CPTAC network on data integration and bioinformatics analysis, as well as translational applications.