Proteomics

蛋白质组学

• 批准号: 8350782
• 负责人: PAUL M STEMMER
• 金额: $ 4.26万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-06 至 2015-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8350782
• 关键词: Algorithms; Amino Acid Sequence; Archives; Area; Binding; Bioinformatics; Biological Assay; Biological Markers; Body Fluids; Cancer Detection; Cell physiology; Cells; Collection; Complex Mixtures; Computational Biology; Computer software; Data; Data Analyses; Data Quality; Deposition; Detection; Digestion; Disease; Dissociation; Educational process of instructing; Electron Transport; Equipment; Event; Experimental Designs; Fluorescence Polarization; Future; Gel; High Pressure Liquid Chromatography; Human Resources; Immunoprecipitation; Individual; Infusion procedures; Instruction; International; Ions; Isoelectric Focusing; Label; Liquid substance; Malignant Neoplasms; Manuals; Mass Spectrum Analysis; Measurement; Methods; Mission; Modification; Molecular Biology; Molecular Genetics; Monitor; Pathway Analysis; Phase; Phosphopeptides; Phosphorylation; Post-Translational Protein Processing; Preparation; Procedures; Process; Productivity; Protein Binding; Protein Chemistry; Proteins; Proteome; Proteomics; Protocols documentation; Published Comment; Reagent; Relative (related person); Research; Research Personnel; Resource Sharing; Resources; Robot; Robotics; Running; Sampling; Savings; Services; Signal Pathway; Signal Transduction; Silver; Solid; Source; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Surface Plasmon Resonance; System; Techniques; Technology; Testing; Therapeutic; Time; Training; Training Support; Treatment Effectiveness; United States National Institutes of Health; Validation; Walking; Western Blotting; anticancer research; base; biological systems; clinically relevant; cost; improved; instrument; instrumentation; liquid chromatography mass spectrometry; mass spectrometer; meetings; member; metaplastic cell transformation; multiple reaction monitoring; nano; programs; protein function; protein profiling; protein protein interaction; scaffold; software systems

The Proteomics Core enhances the research productivity of KCI members by providing the equipment and trained personnel necessary for analysis of cellular protein composition, protein modification, protein quantitation and protein interaction. Proteome profiling and protein identification services utilize modern mass spectrometer based methods. The primary platform for analysis is the Thermo Finnigan LTQ Linear lon Trap equipped with Electron Transfer Dissociation (ETD). Isolated protein, gel plug and full proteome analysis are supported. Sample preparation is achieved by robotic or manual depletion of high abundance proteins, digestion and solid phase extraction (SPE). Various sorbents including specialized sorbents such as Ti02 for isolation of phosphopeptides are available for SPE. Nanoflow HPLC from a Michrom H4 platform is utilized for most analyses with a Triversa Nanomate robot available as needed. Data analysis is achieved using Mascot, Sequest, XITandem and PEAKS algorithms with secondary data analysis by Scaffold. Results are distributed as hard copy on CD or by deposition on the international Tranche network. The Core enhances research productivity by providing a clear and easily accessible mechanism for protein identification and for relative quantitation of proteins based on isobaric tags. Quantitation technologies supported include cICAT, ITraq, TMT and SILAC and Multiple Reaction Monitoring (MRM). Analysis of isotopically labeled samples is achieved using the Mascot Quantitation package. MRM analysis is achieved using the TSQ Vantage with PinPoint and Skyline software for experimental design and data analysis. The protein identification component of the Proteomics Core provides KCI members access to technology for protein identification, proteomic profiling and biomarker identification. The protein interactions component of the Core provides instrumentation and services for detection of protein binding by Fluorescence Polarization (FP) and Surface Plasmon Resonance (SPR). The instruments in the Core produce sensitive, accurate and real time measurements of protein binding events. Thus, the protein interactions component of the Core supports investigators in asking questions about protein-protein interactions and the effects of those interactions on signaling pathways and cellular function.

蛋白质组学核心通过提供设备和 分析细胞蛋白质组成、蛋白质修饰、 定量和蛋白质相互作用。蛋白质组分析和蛋白质鉴定服务利用现代 基于质谱仪的方法。主要分析平台是Thermo Finnigan LTQ Linear 离子阱配备电子转移解离（ETD）。分离蛋白质、凝胶塞和完整蛋白质组 分析得到支持。样品制备是通过机器人或手动消耗高丰度 蛋白质，消化和固相萃取（SPE）。各种吸附剂，包括专用吸附剂， 因为用于分离磷酸肽的TiO 2可用于SPE。来自Michrom H4的Nanoflow HPLC 平台用于大多数分析，并根据需要提供Triversa Nanomate机器人。数据分析是 使用Mascot、Sequest、XITandem和PEAKS算法进行二次数据分析， 脚手架研究结果以光盘硬拷贝或存放在国际Tranche网络上的方式分发。 核心通过为蛋白质提供清晰且易于获得的机制来提高研究生产力 用于基于同量异位素标签的蛋白质的鉴定和相对定量。定量技术 支持的项目包括cICAT、ITraq、TMT和SILAC以及多反应监测（MRM）。分析 使用Mascot定量包获得同位素标记的样品。实现了MRM分析 使用带有PinPoint和Skyline软件的TSQ Vantage进行实验设计和数据分析。的 蛋白质组学核心的蛋白质鉴定部分为KCI成员提供了技术， 蛋白质鉴定、蛋白质组学分析和生物标志物鉴定。 核心的蛋白质相互作用组件提供了检测蛋白质相互作用的仪器和服务。 通过荧光偏振（FP）和表面等离子体共振（SPR）检测蛋白结合。仪器 在核心产生灵敏，准确和真实的时间测量蛋白质结合事件。因此 核心的蛋白质相互作用部分支持研究人员提出有关蛋白质-蛋白质相互作用的问题 相互作用以及这些相互作用对信号传导途径和细胞功能的影响。