COBRE: WVU: SIGNAL TRANSDUCTION & CANCER: MASS SPECTROMETRY & PROTEOMICS CORE

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。WVU蛋白质组学核心设施成立于2001年，是COBRE资助的一部分，目前为WVU和邻近机构提供MALDI-TOF-MS和LC-MS/MS蛋白质鉴定和二维凝胶蛋白分离和分析服务。该核心将有五个功能：1)两种不同样品（健康与患病）之间的差异表达分析或鉴定和相对定量的上调和下调蛋白。2)功能蛋白分析，分离和鉴定复合物中相关的蛋白。3)蛋白质鉴定，对凝胶电泳、western blotting或色谱分离的蛋白质进行鉴定。4)微表征，这将识别和绘制蛋白质的翻译后修饰。5)结构分析，利用距离约束来确定蛋白质的三维结构。这个最终的能力包括交联蛋白质在特定的官能团，蛋白质水解消化和质谱分析。交联肽的质谱分析提供了这些肽在三维结构中的接近性信息。体现在第一种功能中的可以是测定细胞、组织或生物液体的蛋白质组补体。这份更新申请描述了：1)该设施的历史，目前的仪器和人员，2)核心设施运营计划，3)核心设施在实现COBRE资助的研究目标和WVU其他研究项目中的作用，4)在COBRE资助的下一个资助周期中将由核心设施支持的信号转导和癌症研究项目，以及5)为该设施获得额外资金的努力进展。

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. The WVU Proteomics Core Facility was established in 2001 as part of the COBRE grant, and is currently offering MALDI-TOF-MS and LC-MS/MS protein identification and 2-D gel protein separations and analysis as a service to WVU and neighboring institutions. This core will serve five functions: 1) Differential expression analysis or the identification and relative quantification of up- and down-regulated proteins between two different samples (health versus diseased). 2) Functional protein analysis, which will isolate and identify proteins associated in complexes. 3) Protein identification, which will identify proteins isolated by gel electrophoresis, western blotting or chromatography. 4) Micro-characterization, which will identify and map post-translational modifications of proteins. 5) Structural analysis, which will use the distance constraints for determination of a protein¿s three-dimensional structure. This final capability involves cross-linking proteins at specific functional groups, proteolytic digestion, and MS analysis. MS analysis of cross-linked peptides provides information on proximity of these peptides in three-dimensional structure. Embodied in the first function can be the determination of the proteomic complement of a cell, tissue or biological fluid. This renewal application describes: 1) the Facility¿s history, current instrumentation, and personnel, 2) the Core Facility Operational Plan, 3) the role of the Core Facility in fulfilling research objectives in COBRE-funded and other research programs at WVU, 4) research projects in ¿Signal Transduction and Cancer¿ that will be supported by the Core Facility in the next funding cycle of the COBRE grant, and 5) progress on efforts to obtain additional funding for the Facility.