Supplement for Cobre in Signal Transduction and Cancer R

描述（由申请人提供）： 信号转导和癌症COBRE（RR 16440）在西弗吉尼亚大学建立了蛋白质组学设施。该大学捐赠了900平方英尺的翻新实验室空间。一台LC-MS质谱仪被借给蛋白质组学核心设施，以启动该核心，而一台较旧的Millipore大型2-D设备被捐赠给该设施。Cobre为小型设备提供了资金，如拔针器、凝胶设备和离心机。此外，Cobre还为支持人员提供资金，包括核心设施主任和一名技术员/质谱专家。随后，NCRR（S 10-RR 16792 -01）资助了一笔设备赠款，为该设施提供了两台专用质谱仪，一台Maldi-TOF和一台LC-MS离子阱质谱仪。癌症中心和医学院已承诺长期支持该设施，并成立了一个指导委员会。然而，为了对癌细胞和组织进行蛋白质组学分析，核心设施需要额外的蛋白质组学相关设备，使研究人员能够从2-D凝胶中识别感兴趣的蛋白质“点”，2-D凝胶的图像和目录数据库，分离单个蛋白质“点”，消化它们并为序列分析做好准备。执行这些功能的设备将有助于蛋白质组学设施的自动化，并将与Maldi的高通量能力和LC-MS离子阱质谱的蛋白质测序能力无缝连接。在本Cobre补充请求中，我们寻求捆绑的主要蛋白质组学设备和软件，以执行这些功能。目的1：蛋白质纯化设备，包括HPLC和Millipore水纯化。目的2：一种新型的蛋白质双向凝胶分离装置.目标3：扫描仪，用于二维凝胶成像，识别感兴趣的蛋白质“点”并生成数据库，目标4：点拾取器，其具有与扫描仪集成的相关软件和可以从凝胶或PVDF拾取单个蛋白质点的机械臂。目的5：一个消化系统，允许胰蛋白酶化和洗脱个别蛋白质。目标6：计算机为数据分析提供工作站。总的来说，这套捆绑设备将完成我们的蛋白质组学设施。这些数据将使我们的癌症研究科学家能够识别新的蛋白质靶点和在肿瘤细胞系和组织中特异性激活的蛋白质。我们假设这些信息将使研究人员能够帮助医生根据在个体肿瘤活检中发现的独特的信号蛋白阵列为个体定制癌症治疗。

DESCRIPTION (provided by applicant): The COBRE for Signal Transduction and Cancer (RR16440) established a Proteomics Facility at West Virginia University. The University donated 900 r2 of renovated lab space. A LC-MS mass spectrometers was loaned to the Proteomics core facility, to initiate this core, while an older Millipore large format 2-D apparatus was donated to the facility. Cobre funds provided for small equipment such as a needle puller, gel equipment and centrifuges. In addition, Cobre funds support personnel, including the core facility director and a technician/mass spectrometry specialist. Subsequently, an equipment grant was funded by the NCRR (S 10-RR16792-01) that provided two dedicated mass spectrometers for the facility, a Maldi-TOF and an LC-MS ion trap mass spectrometer. The cancer center and school of medicine have pledged long term support for the facility, and a steering committee has been established. However, in order to perform proteomic analysis on cancer cells and tissues, the core facility requires additional proteomic-related equipment that enable researchers to identify protein "spots" of interest from 2-D gels, image and catalog databases of 2-D gels, isolate the individual protein "spots", digest them and prepare them for sequence analysis. Equipment that perform these functions would help to automate the Proteomics Facility and would seamlessly link with the high throughput capabilities of Maldi and the protein sequencing capabilities of LC-MS ion trap mass spectrometry. In this Cobre supplemental request, we seek bundled major proteomic equipment and software that perform these functions. Aim 1: protein purification equipment including HPLC and Millipore water purification. Aim 2: a newer 2-D gel apparatus protein separation. Aim 3: a scanner, to image 2-D gels, identify protein "spots" of interest and generate databases, Aim 4: a spot picker, which has associated software that is integrated with the scanner and a robotic arm that can pick individual protein spots from a gel or PVDF. Aim 5: a digestion system that permits tryptsinazation and elution of individual proteins. Aim 6: computers to provide work stations for data analysis. Collectively, this set of bundled equipment will complete our Proteomics facility. These data will enable our cancer research scientists to identify novel protein targets and proteins that are specifically activated in tumor cell lines and tissues. We hypothesize that this information will enable researchers to assist physicians in customizing cancer treatment to individuals, based on the unique array of signaling proteins found to be activated in an individual tumor biopsy.