Proteomics Core

蛋白质组学核心

• 批准号: 8374855
• 负责人: DAVID L HACHEY
• 金额: $ 21.51万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8374855
• 关键词: Address; Affinity; Affinity Chromatography; Algorithms; Aliquot; Area; Blood capillaries; Cations; Cell Lineage; Cell physiology; Charge; Chemicals; Collection; Complement; Complex; Complex Mixtures; Computer software; Consultations; Coupled; Critiques; Custom; Data; Data Analyses; Data Set; Databases; Detection; Deuterium; Development; Digestion; Disease Marker; Education; Electrospray Ionization; Ensure; Enzymes; Epithelial Cells; Event; Excision; Experimental Designs; Fluorescent Dyes; Gel; Genome; Global Change; Helicobacter pylori; High Pressure Liquid Chromatography; Human Resources; Image; Individual; Inflammation; Informatics; Investigation; Ion Exchange; Ions; Isotopically-Coded Affinity Tagging; Label; Laboratories; Lead; Linux; Location; MALDI-TOF Mass Spectrometry; Maleimides; Maps; Mass Spectrum Analysis; Measures; Metals; Methodology; Methods; Mining; Minor; Modification; Molecular Analysis; Monitor; Multiprotein Complexes; Nature; Pattern; Peptide Fragments; Peptide Hydrolases; Peptide Mapping; Peptides; Phase; Phosphopeptides; Phosphorylation; Phosphotyrosine; Post-Translational Protein Processing; Postdoctoral Fellow; Preparation; Process; Program Research Project Grants; Protein Isoforms; Proteins; Proteolysis; Proteome; Proteomics; Reaction; Reagent; Relative (related person); Research; Research Personnel; Research Project Grants; Resources; Running; Sampling; Scanning; Services; Signal Transduction; Sodium Chloride; Software Tools; Solutions; Source; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Speed; Spottings; Stable Isotope Labeling; Staging; Staining method; Stains; Stomach; Students; System; Techniques; Technology; Therapeutic; Training; Trypsin; Variant; Work; adduct; base; biological systems; capillary; comparative; cyanine dye; cyanine dye 5; data mining; database design; design; flexibility; follow-up; gel electrophoresis; instrument; instrumentation; intercellular communication; interest; ionization; mRNA Differential Displays; malignant stomach neoplasm; mass spectrometer; migration; multicore processor; nanolitre; new technology; novel; novel strategies; open source; protein expression; research study; response; software development; stable isotope; stoichiometry; tandem mass spectrometry; therapeutic target; tool

The Proteomics Core (Core B) provides state-of-the-art instrumentation and expertise in analytical proteomics to investigators in this PPG. A unique feature of the Proteomics Core is the availability of all major proteomics analysis technologies, which offers exceptional flexibility in analytical approaches. The Proteomics Core provides 1) identification of proteins in both simple and complex multiprotein samples, ncluding protein components of multiprotein complexes, 2) quantitative analyses of differential protein expression and modification, and 3) analyses of protein modifications. Proteomics Core staff provide consultation on experimental design and sample preparation, as well as digestion, protein and peptide separations, MS analyses and primary data workup. Both electrospray onization (ESI) mass spectrometers for LC-MS-MS experiments and MALDI TOF-TOF instruments are available for MS analyses. DIGE/MS technology includes fluorescent dyes and multiplexed 2D gels with nternal standards for direct quantitative comparisons. Automated workstations are used for protein excision and sample processing, followed by MALDI-TOF MS and TOF/TOF MS/MS and automated database searching with Mascot software for protein identification. The Core also offers relative quantitation with Nterminal reactive differential labeling reagents (iTRAQ and isotopically-labeled phenylisocyanate) for relative quantitation of modified protein forms. For protein identification from multiprotein complexes, multidimensional LC is coupled directly with automated LC-MS-MS, followed by automated database searching with Sequest and Mascot software. Sequest searches are done on a high speed, multiprocessor Linux cluster in the Advanced Computing Center for Research and Education (ACCRE). Sequest-based identifications are evaluated and filtered using open-source tools (Peptide/Protein Prophet) and integrated with a custom-designed database system (CHIPS) for organizing, filtering and comparing data sets. Additional data analysis by SALSA and P-Mod enables identification of posttranslational modifications, adducts or sequence variants from MS-MS data. Project 1 (Peek) will utilize the DIGE/MS differential-display technology, and Project 3 (Cover) will utilize the protein identification technologies from both simple and complex multiprotein samples. Project 2 (Polk) will heavily utilize both services. Analysis of protein modifications is no longer proposed in the revised Projects, but may be used for subsequent/complementary studies.

蛋白质组学核心（核心B）提供了最先进的仪器和分析方面的专业知识 蛋白质组学向研究人员中的蛋白质组学。蛋白质组学核心的独特功能是所有人的可用性 主要的蛋白质组学分析技术，在分析方法中提供了出色的灵活性。这 蛋白质组学核心1）在简单和复杂的多蛋白样品中鉴定蛋白质， 多蛋白质复合物的NCLUD蛋白成分，2）差异蛋白的定量分析 表达和修饰，以及3）蛋白质修饰的分析。 蛋白质组学核心人员提供有关实验设计和样品准备的咨询，以及 消化，蛋白质和肽分离，MS分析和主要数据工作。两个电喷雾 LC-MS-MS实验和MALDI TOF-TOF仪器的ONIzation（ESI）质谱仪是 可用于MS分析。 DIGE/MS技术包括荧光染料和多重2D凝胶 直接定量比较的NENTER标准。自动化工作站用于蛋白质切除 和样品处理，然后是MALDI-TOF MS和TOF/TOF/TOF MS/MS和自动数据库 使用吉祥物软件搜索蛋白质识别。核心还提供相对定量的NTerminal 相对的反应性差异标记试剂（ITRAQ和同位素标记的苯糖氰酸酯） 定量改性蛋白质形式。对于来自多蛋白复合物的蛋白质鉴定， 多维LC与自动LC-MS-MS直接耦合，然后是自动数据库 使用Secest和Mascot软件进行搜索。隔离搜索是在高速，多处理器上进行的 Linux群集在高级研究与教育中心（ACP）中。基于续集 使用开源工具（肽/蛋白质先知）评估并过滤识别，并集成 使用定制设计的数据库系统（芯片），用于组织，过滤和比较数据集。 Salsa和P-Mod的其他数据分析可以鉴定翻译后修改， 来自MS-MS数据的加合物或序列变体。 项目1（PEEK）将利用DIGE/MS差分播放技术，项目3（封面）将 利用简单和复杂多蛋白样品的蛋白质识别技术。项目2 （Polk）将大量利用这两种服务。在修订后不再提出蛋白质修饰的分析 项目，但可用于随后/互补研究。