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

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 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。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 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.