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

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子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相关的发展，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。 本年度报告提供了中心研究人员自上而下的蛋白质组学进展;描述了FAIMS应用气相分馏的进展;翻译后修饰的测量，包括磷酸化蛋白质组，除了更丰富的pSer和pThr修饰外，还测量了低丰度pTyr修饰;报告了RePlay蛋白质组学的增强版本。 先进的样品制备方法（例如，胰蛋白酶包被的纳米颗粒;生物流体CSF和唾液）、压力消化和超声处理，已经被开发用于支持测量用于系统生物学研究的差异蛋白质丰度。 核心1还包括基于活性的蛋白质组学（ABP和MABPP）的研究领域，提供了一种更有针对性的方法来探测组织，细胞，亚细胞组分和生物流体的亚蛋白质组。 该中心正在实施亚细胞分离方法，以实现核心1的目标，即表征细胞中的蛋白质及其相关的定位和运输。 在细胞表面膜蛋白质组、精子成熟、致癌病毒检测和基因组注释方面也取得了新的进展。

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.