Proteomics

蛋白质组学

基本信息

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
项目类别：
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

项目摘要

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，蛋白质所需的训练有素的人员定量和蛋白质相互作用。蛋白质组分析和蛋白质识别服务使用现代基于质谱仪的方法。分析的主要平台是Thermo Finnigan LTQ线性配备电子传递解离（ETD）的LON陷阱。孤立的蛋白质，凝胶塞和全蛋白质组支持分析。样品制备是通过机器人或手动耗尽高丰度来实现的蛋白质，消化和固相提取（SPE）。各种吸附剂，包括专业吸附剂由于用于分离磷酸肽的Ti02可用于SPE。 MICHROM H4的Nanoflow HPLC 使用Triversa纳米酸盐机器人可用，用于大多数分析的平台。数据分析是使用吉祥物，序列，Xitandem和峰算法实现了辅助数据分析脚手架。结果在CD上的硬拷贝或通过沉积在国际批量网络上分布。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率鉴定和基于同类标签的蛋白质相对定量。定量技术支持包括CICAT，ITRAQ，TMT和SILAC以及多重反应监测（MRM）。分析使用吉祥物定量软件包实现了同位素标记的样品。实现MRM分析将TSQ Vantage与Pinpoint和Skyline软件一起进行实验设计和数据分析。这蛋白质组学核心的蛋白质识别成分为KCI成员提供了对技术的访问蛋白质鉴定，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部分提供了仪器和服务以检测通过荧光极化（FP）和表面等离子体共振（SPR）结合蛋白质。乐器在核心中，蛋白质结合事件产生敏感，准确和实时测量。因此，核心的蛋白质相互作用组成部分支持研究人员询问有关蛋白质蛋白质的问题相互作用以及这些相互作用对信号通路和细胞功能的影响。