Supplement for Cobre in Signal Transduction and Cancer R

描述(由申请人提供)： 信号转导和癌症中心(RR16440)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个R2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，以启动这个核心设施，而一台更老的米利波尔大幅面2-D仪器被捐赠给该设施。科布雷基金为小型设备提供资金，如拉针器、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人员/质谱仪专家。随后，国家科研院(S 10-RR16792-01)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离子陷阱质谱仪。癌症中心和医学院已经承诺为该设施提供长期支持，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分析，核心设施需要额外的蛋白质组相关设备，使研究人员能够从二维凝胶、二维凝胶的图像和目录数据库中识别感兴趣的蛋白质“点”，分离单个蛋白质“点”，消化它们并准备进行序列分析。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的离子陷阱质谱仪的蛋白质测序能力无缝连接。在这份Cobre补充申请中，我们寻求执行这些功能的捆绑主要蛋白质组设备和软件。目的1：蛋白质纯化设备，包括高效液相色谱法和微孔水净化。目的2：一种新型的二维凝胶蛋白质分离装置。目标3：一种扫描仪，用于对2-D凝胶成像，识别感兴趣的蛋白质“斑点”并生成数据库，目标4：斑点拾取器，它具有与扫描仪集成的相关软件，以及可以从凝胶或PVDF中挑选单个蛋白质斑点的机械臂。目标5：一种允许对单个蛋白质进行胰酶消化和洗脱的消化系统。目标6：计算机为数据分析提供工作站。总的来说，这套捆绑的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标和在肿瘤细胞系和组织中特异激活的蛋白质。我们假设，这些信息将使研究人员能够帮助医生根据个人肿瘤活检中发现激活的独特信号蛋白阵列，定制针对个人的癌症治疗。

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.