Proteomics and Metabolomics

蛋白质组学和代谢组学

• 批准号: 8180857
• 负责人: Louis M. Weiner
• 金额: $ 10.11万
• 依托单位: GEORGETOWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-06 至 2013-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8180857
• 关键词: Proteomics; Metabolomics

Proteomics and Metabolomics Shared Resource-Stephen Byers, PhD /Albert Fornace, Jr, MD The mission of the Proteomics and Metabolomics Shared Resource (PMSR) is to provide state-of-the-art tools, methods, and mass spectrometry-based technology to investigators. This centralized shared resource is available to researchers for routine and specialized proteomic and metabolomic applications. The aim of PMSR is to provide an 'in house' resource, rich in technical expertise that fosters the free flow of experimental knowledge and collaborations across the Lombardi Comprehensive Cancer Center (Lombardi). The PMSR offers an array of proteomics- and metabolomics-based services. It is equipped with two-dimensional (2D) gel electrophoresis apparatus including DIGE imager and spot picker (GE Healthcare), 4800 MALDI TOF-TOF instrument (ABI), and a QSTAR Elite Hybrid LC MS/MS system (ABI) to support the proteomic workflow. Each instrument has the latest integrated software and database search engines (e.g.. Protein Pilot, GPS explorer) for protein identification and data processing. In addition, PMSR houses a QTOF Premier (Waters Corp.) online with an UPLC system for metabolomic profiling and drug metabolism studies. The data processing for metabolite biomarker studies is supported by the 'SIMCA-P' and the 'Random Forest software'. PMSR also manages a 4000 QTrap, which supports quantitation and validation for small-molecule metabolites, and allows targeted proteomic analysis. PMSR staff includes a dedicated bioinformatician who works closely with the Biostatistics and Bioinformatics Shared Resource (BBSR), as well as with the Georgetown University-based Protein Information Resource (PIR). The most common experiments include identification of proteins contained in bands/spots isolated from electrophoretic gels, as well as comparative proteomics using shotgun and stable isotope labeling strategies. The high speed, sensitivity, and accuracy of our mass spectrometry experiments allow accurate qualitative and quantitative proteomics. The metabolomics initiative has been driven by Dr. Albert Fornace, Jr, and involves UPLC-TOFMS based biomarker discovery for radiation exposure. This technology is now available to investigators for their specific research interests. Written and electronic reports are provided for all samples, and the average turn-around time is two weeks. The PMSR is able to carry out experiments that detect and characterize protein modifications such as phosphorylation and acetylafion. These experiments are challenging and require a more intensive effort. However, the combination of the complementary mass spectrometric methods available in PMSR, staff expertise and experience, and the ability to work closely with investigators, have allowed the successful completion of a number of these difficult projects. The PMSR was established in January 2006 and has grown dramatically, both in resources and use. Lombardi has invested over $1 million in equipment alone for the PMSR; as a result, the facility consistently provides high quality data to many investigators from 5 programs within Lombardi. The PMSR is run under the directorship of Drs. Stephen Byers and Albert Fornace Jr.

蛋白质组学和代谢组学共享资源-Stephen Byers 博士/Albert Fornace Jr 医学博士 蛋白质组学和代谢组学共享资源 (PMSR) 的使命是为研究人员提供最先进的工具、方法和基于质谱的技术。这种集中的共享资源可供研究人员用于常规和专门的蛋白质组学和代谢组学应用。 PMSR 的目标是提供“内部”资源，拥有丰富的技术专业知识，促进实验的自由流动 整个隆巴迪综合癌症中心（Lombardi）的知识和合作。 PMSR 提供​​一系列基于蛋白质组学和代谢组学的服务。它配备了二维 (2D) 凝胶电泳设备，包括 DIGE 成像仪和点选择器 (GE Healthcare)、4800 MALDI TOF-TOF 仪器 (ABI) 和 QSTAR Elite Hybrid LC MS/MS 系统 (ABI)，以支持蛋白质组工作流程。每个 仪器拥有最新的集成软件和数据库搜索引擎（例如 Protein Pilot、GPS explorer），用于蛋白质鉴定和数据处理。此外，PMSR 还配备了 QTOF Premier（沃特世公司）和 UPLC 系统，用于代谢组学分析和药物代谢研究。代谢物生物标志物研究的数据处理由“SIMCA-P”和“随机森林软件”支持。永磁同步谐振器也 管理 4000 QTrap，支持小分子代谢物的定量和验证，并允许进行有针对性的蛋白质组分析。 PMSR 工作人员包括一名专门的生物信息学家，他与生物统计学和生物信息学共享资源 (BBSR) 以及乔治城大学蛋白质信息资源 (PIR) 密切合作。最常见的实验包括鉴定所含蛋白质 从电泳凝胶中分离出的条带/斑点，以及使用鸟枪法和稳定同位素标记策略的比较蛋白质组学。我们的质谱实验速度快、灵敏度高且准确，可实现准确的定性和定量蛋白质组学。该代谢组学计划由 Albert Fornace, Jr. 博士推动，涉及基于 UPLC-TOFMS 的辐射暴露生物标志物发现。现在，研究人员可以根据他们的特定研究兴趣使用这项技术。书面和电子报告 所有样品均提供，平均周转时间为两周。 PMSR 能够进行检测和表征蛋白质修饰（例如磷酸化和乙酰化）的实验。这些实验具有挑战性，需要付出更大的努力。然而，PMSR 中可用的补充质谱方法、工作人员的专业知识和经验以及与研究人员密切合作的能力相结合，使得许多这些困难的项目得以成功完成。 PMSR 成立于 2006 年 1 月，在资源和使用方面都取得了显着增长。 Lombardi 仅在 PMSR 设备上就投资了超过 100 万美元；因此，该设施始终为 Lombardi 内 5 个项目的许多研究人员提供高质量的数据。 PMSR 在博士的领导下运行。斯蒂芬·拜尔斯和小阿尔伯特·福纳斯