APPROACHES FOR PROTEIN MODIFICATIONS, INTERACTIONS, & SPATIAL & QUANTITATIVE DYN

蛋白质修饰、相互作用的方法，

• 批准号: 8365459
• 负责人: RICHARD D SMITH
• 金额: $ 34.41万
• 依托单位: BATTELLE PACIFIC NORTHWEST LABORATORIES
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8365459
• 关键词: Annual Reports; Area; Biology; Cell Fractionation; Cell surface; Cells; Development; Digestion; Fractionation; Funding; Gases; Genome; Goals; Grant; Measurement; Measures; Membrane; Methods; Modification; National Center for Research Resources; Oncogenic Viruses; Phase; Post-Translational Protein Processing; Preparation; Principal Investigator; Proteins; Proteome; Proteomics; Reporting; Research; Research Infrastructure; Research Personnel; Resources; Saliva; Sampling; Sonication; Source; Sperm Maturation; Subcellular Fractions; Systems Biology; Tissues; Trypsin; United States National Institutes of Health; base; cost; nanoparticle; pressure; protein protein interaction; protein transport; trafficking; viral detection

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Core 1 focuses on developments associated protein localization within the cell, the protein-protein interactions, the quantitative dynamics and protein trafficking, and the modification states of proteins applying high throughput proteomics measurements. For this annual report advancements in top-down proteomics by Center-investigators are provided; advancement in applying gas phase fractionation with FAIMS are described; measurements of posttranslational modifications including the phosphoproteome with measurements of low abundance pTyr modifications in addition to the more abundant pSer and pThr modifications; and an enhanced version of RePlay proteomics is reported. Advanced methods of sample preparation (e.g., trypsin coated nanoparticles; the biofluids CSF and saliva), pressure digestion and sonication, have been developed in support of measuring differential protein abundances for systems biology studies. Core 1 also includes the research area of activity-based proteomics (ABP and MABPP) that provides a more targeted approach to probing the sub-proteomes of tissues, cells, subcellular fractions, and biofluids. Methods in subcellular fractionation are being implemented in the Center to achieve Core 1 goals for characterizing proteins in cells and their associated localization and trafficking. Additional progress has been made on profiling the cell surface membrane proteome, sperm maturation, oncogenic virus detection, and genome annotation.

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 核心1侧重于与细胞内蛋白质定位、蛋白质-蛋白质相互作用、定量动力学和蛋白质运输以及应用高通量蛋白质组学测量的蛋白质修饰状态相关的研究进展。 在这份年度报告中，提供了中心研究人员在自上而下蛋白质组学方面的进展；描述了用FAIMS应用气相分离的进展；翻译后修饰的测量，包括低丰度pTyr修饰的测量，以及更丰富的pSer和pThr修饰；以及重放蛋白质组学的增强版本的报告。已开发出先进的样品制备方法(例如，胰酶包被的纳米颗粒；生物流体脑脊液和唾液)、加压消化和超声波处理，以支持系统生物学研究测量差异蛋白质丰度。核心1还包括基于活性的蛋白质组学(ABP和MABPP)的研究领域，该领域提供了一种更有针对性的方法来探测组织、细胞、亚细胞部分和生物液的亚蛋白质组。该中心正在实施亚细胞分离的方法，以实现确定细胞中蛋白质及其相关的定位和运输的核心1目标。在细胞表膜蛋白质组、精子成熟、致癌病毒检测和基因组注释方面取得了更多进展。