Novel Quantitative, Comprehensive Proteomic Technology

新颖的定量、综合蛋白质组学技术

• 批准号: 6666673
• 负责人: Ruedi H. Aebersold
• 金额: $ 56.38万
• 依托单位: INSTITUTE FOR SYSTEMS BIOLOGY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-28 至 2005-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6666673
• 关键词: affinity chromatography; chemical information system; computer system design /evaluation; hormone sensitivity /resistance; informatics; mass spectrometry; oncoproteins; prostate neoplasms; proteins; proteomics; technology /technique development

DESCRIPTION: (provided by applicant) Global genome initiatives have generated enormous amounts of information, spurned new technologies and catalyzed the emergence of a new type of biology which attempts to build biological knowledge from systematic and quantitative analysis of the molecules that constitute a cell or tissue. Expectations for applying comprehensive and quantitative analyses of genes and proteins to cancer biology are high. Comparative profiling of mRNA & proteins expressed in "normal" and cancer cells and tissues is expected to discover new markers for diagnosis of type, stage and stratification of cancer, indicators to assess success of treatment and emergence of drug resistance, and lead to an understanding of the molecular basis of cancer and thus new therapeutic targets. We propose to develop a novel technology for quantitative, sensitive and comprehensive analysis of protein expression profiles in human cells and tissues. It is based on quantitative, automated mass spectrometry of complex peptide mixtures and consists of 2 phases. First is generation of a reference peptide database of a given cell type in which each peptide is annotated with characteristics that uniquely identify it in the database. Second is correlation of mass spectrometric data obtained from clinical or research samples with the peptide database to identify and quantify protein components of the cell type. The technology is: 1) automatable, 2) compatible with analysis of essentially any protein, 3) robust and easy to standardize and 4) amenable to high throughput. This technology will provide a general/robust technical basis for comprehensive and quantitative analysis of protein profiles of normal and cancer cells and tissues that will be widely accessible. The new technology will be validated and applied to prostate cancer research. Specifically, we will correlate the data obtained from the quantitative protein profiling technology developed in this program with gene expression data obtained via cDNA arrays and Lynx Therapeutic's massively parallel signature sequencing (MPSS) procedure to identify biologic determinants that differentiate indolent and aggressive prostate cancer and to determine the molecular basis of androgen dependent cell growth of prostate cells. The application satisfies the request for innovative technology development in 3 ways: First, the technology explores a new approach to identification and quantification of the comprehensive spectrum of proteins expressed in human cells/tissues; Second, the annotated peptide database (APD) for prostate tissue will be made available publicly and serve as a necessary reference for quantitative protein expression profiling in laboratories not currently specialized in proteomics and Third, research will be conducted as an academic/industrial partnership assuring rapid commercial accessibility of reagents/instrumentation supporting the technology. We anticipate the proposed pro ram will contribute significant towards fulfilling expectations for cancer research raised by the genomics revolution.

描述：（由申请人提供） 全球基因组倡议产生了大量信息， 拒绝新技术并催化新型生物学的出现 它试图从系统和定量的角度建立生物学知识 分析构成细胞或组织的分子。对的期望 应用基因和蛋白质的全面定量分析 癌症生物学水平很高。 mRNA 和蛋白质表达的比较分析 “正常”细胞和癌细胞和组织有望发现新的标记物 癌症类型、分期和分层的诊断、评估指标 治疗的成功和耐药性的出现，并导致 了解癌症的分子基础以及新的治疗方法 目标。我们建议开发一种定量、灵敏的新技术 以及人体细胞中蛋白质表达谱的综合分析 组织。它基于复杂物质的定量、自动质谱分析 肽混合物由两相组成。首先是生成参考 给定细胞类型的肽数据库，其中每个肽都注释为 在数据库中唯一标识它的特征。其次是 从临床或研究获得的质谱数据的相关性 使用肽数据库来识别和量化蛋白质成分的样品 的细胞类型。该技术是：1) 可自动化，2) 兼容 几乎可以分析任何蛋白质，3) 稳健且易于标准化，4) 适合高吞吐量。该技术将提供通用/鲁棒的 蛋白质综合定量分析技术基础 正常细胞和癌细胞和组织的概况将被广泛应用 可以访问。新技术将被验证并应用于前列腺 癌症研究。具体来说，我们将关联从 本项目开发的基因定量蛋白质分析技术 通过 cDNA 阵列和 Lynx Therapeutic 大规模获得的表达数据 并行特征测序 (MPSS) 程序来识别生物制品 区分惰性前列腺癌和侵袭性前列腺癌的决定因素 确定前列腺雄激素依赖性细胞生长的分子基础 细胞。该应用满足创新技术的要求 发展有3个方面：一是技术探索新途径 蛋白质综合谱的鉴定和定量 在人体细胞/组织中表达；二、注释肽数据库（APD） 前列腺组织将公开提供并作为必要的 实验室定量蛋白质表达谱的参考不 目前专门从事蛋白质组学，第三，研究将作为 学术/工业合作伙伴关系确保快速商业化 支持该技术的试剂/仪器。我们预计拟议的 Pro RAM 将为满足癌症期望做出重大贡献 基因组学革命引发的研究。

项目成果

LC-MS solvent composition monitoring and chromatography alignment using mobile phase tracer molecules.

使用流动相示踪分子进行 LC-MS 溶剂成分监测和色谱比对。

• DOI: 10.1016/j.jchromb.2005.09.039
• 发表时间: 2005
• 期刊: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences
• 作者: Chen,SharonS;Aebersold,Ruedi
• 通讯作者: Aebersold,Ruedi

Identification of androgen-coregulated protein networks from the microsomes of human prostate cancer cells.

• DOI: 10.1186/gb-2003-5-1-r4
• 发表时间: 2003
• 期刊: Genome biology
• 影响因子: 12.3
• 作者: Wright ME;Eng J;Sherman J;Hockenbery DM;Nelson PS;Galitski T;Aebersold R
• 通讯作者: Aebersold R

Chemical probes and tandem mass spectrometry: a strategy for the quantitative analysis of proteomes and subproteomes.

• DOI: 10.1016/j.cbpa.2003.12.001
• 发表时间: 2004-02
• 期刊: Current opinion in chemical biology
• 影响因子: 7.8
• 作者: Hui Zhang;Wei Yan;R. Aebersold