Core 012 (797) Proteomics Core

项目摘要 蛋白质组学核心（PC）为莫菲特癌症中心（MCC）提供最先进的蛋白质化学服务 成员和尖端的分析技术，以定性和定量评估蛋白质表达， 相互作用、活性、突变和翻译后修饰（PTM）。核心成立于2003年 在2011年的评审中被评为“优秀”可以用细胞系和动物进行实验 模型以及患者标本，包括不同的组织类型和生物液体。这些的范围 研究范围从单个蛋白质的详细分子特征到蛋白质组范围的分析。的 蛋白质组学核心的总体目标是：1）在设计和 执行蛋白质组学实验; 2）提供仪器和训练有素的工作人员来支持成员; （三）对会员及其工作人员进行培训。PC由四名全职工作人员组成， 用于发现蛋白质组学的色谱-串联质谱（LC-MS/MS）肽测序，以及 液相色谱-多反应监测质谱法（LC-MRM）用于靶向定量。 在PC中进行的研究包括检查蛋白质-蛋白质相互作用的研究（使用免疫沉淀法 或串联亲和纯化）、药物靶标（化学蛋白质组学）、信号传导（磷酸蛋白质组学）、蛋白质组- 广泛的乙酰化或泛素化，PTM的顺序富集，以及不同扰动对 选择的蛋白质或蛋白质组（例如，药物治疗、siRNA敲除、CRISPR敲除）。此夕h PC使用代谢标记（SILAC）或化学标记（iTRAQ/TMT）提供样本多重检测 比较蛋白质组学技术。最后，PC使用发现蛋白质组学的集成管道， 探索癌症生物学和目标量化，以实现高度精确的生物标志物测量， 从模型系统转化为患者样本。PC与分子基因组学核心的相互作用 (MGC)是蛋白质基因组学分析的必要条件;佛罗里达东南大学 整合代谢组学中心，支持蛋白质代谢组学。下游与 生物统计学核心（BC）和癌症信息学核心（CIC）有助于理解和评估 复杂蛋白质组学实验的结果。在项目期间，PC支持四个国家和地区的用户， 为47份出版物提供了捐助，比2006-2010年供资期间的15份出版物有所增加。最 在最近一个财政年度，PC支持了26个成员，其中90%的使用量来自同行评审资助的成员。 使用CCSG提供的财政支持和机构资金，PC继续提供国家的- 先进的设备、技术和服务，以满足MCC成员当前和未来的需求。

PROJECT SUMMARY The Proteomics Core (PC) provides state-of-the-art protein chemistry services for Moffitt Cancer Center (MCC) members and cutting-edge analytical techniques to qualitatively and quantitatively assess protein expression, interactions, activity, mutations and post-translational modifications (PTMs). The core was established in 2003 and was scored “Outstanding” in the 2011 review. Experiments can be conducted with cell line and animal models as well as with patient specimens, including diverse tissue types and biofluids. The scope of these studies ranges from detailed molecular characterization of a single protein to proteome-wide profiling. The overall aims of the Proteomics Core are to: 1) consult and collaborate with members in the design and execution of proteomics experiments; 2) provide instrumentation and highly trained staff to support members; and 3) train members and their staff. The PC is comprised of four full-time staff and provides liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide sequencing for discovery proteomics, and liquid chromatography-multiple reaction monitoring mass spectrometry (LC-MRM) for targeted quantification. Studies performed in the PC include those that examine protein-protein interactions (using immunoprecipitation or tandem affinity purification), drug targets (chemical proteomics), signaling (phosphoproteomics), proteome- wide acetylation or ubiquitination, sequential enrichment of PTMs, and the effects of different perturbations on selected proteins or the proteome (e.g., drug treatment, siRNA knockdown, CRISPR knockout). Further, the PC provides sample multiplexing using metabolic labeling (SILAC) or chemical labeling (iTRAQ/TMT) techniques for comparative proteomics. Finally, the PC uses an integrated pipeline of discovery proteomics to explore cancer biology and target quantification for highly precise biomarker measurements that can be translated from model systems into patient specimens. PC interactions with the Molecular Genomics Core (MGC) are necessary for combined proteogenomic analysis; and with the University of Florida South East Center for Integrated Metabolomics, to support proteometabolomics. Downstream interactions with the Biostatistics Core (BC) and the Cancer Informatics Core (CIC) contribute to understanding and evaluating the results of complex proteomics experiments. During the project period, the PC supported users across four and contributed to 47 publications, an increase from 15 publications in the 2006-2010 funding period. In the most recent fiscal year, the PC supported 26 members, with 90% of total usage by peer-review-funded members. Using financial support provided by the CCSG and Institutional funds, the PC continues to provide state-of-the- art equipment, technologies, and services to meet the current and future needs of MCC members.