Core 012 (797) Proteomics Core

项目总结 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)。该中心成立于2003年。 并在2011年的评测中被评为“杰出”。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。这些措施的范围 研究范围从单一蛋白质的详细分子特征到整个蛋白质组的图谱。这个 蛋白质组学核心的总体目标是：1)在设计和开发过程中与成员协商和协作 进行蛋白质组学实验；2)提供仪器和训练有素的工作人员为成员提供支持； 3)培训会员及其工作人员。个人计算机由四名全职工作人员组成，并提供液体 发现蛋白质组学的LC-MS/MS多肽测序，以及 用于靶向定量的液相色谱-多反应监测质谱仪(LC-MRM) 在PC中进行的研究包括那些检查蛋白质-蛋白质相互作用的研究(使用免疫沉淀 或串联亲和纯化)，药物靶标(化学蛋白质组学)，信号(磷蛋白质组学)，蛋白质组- 广泛的乙酰化或泛素化，PTM的顺序浓缩，以及不同扰动对 选定的蛋白质或蛋白质组(例如，药物治疗、siRNA敲除、CRISPR敲除)。此外， PC提供使用代谢标记(SILAC)或化学标记(iTRAQ/TMT)的样本多路传输 比较蛋白质组学技术。最后，PC使用发现蛋白质组学的集成流水线来 探索癌症生物学和目标量化，以获得高精度的生物标记物测量 从模型系统转换成患者样本。PC与分子基因组学核心的相互作用 (MGC)是进行联合蛋白质基因组分析所必需的；并与佛罗里达东南大学合作 综合代谢组学中心，支持蛋白质代谢组学。与下游的交互 生物统计核心(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结果。在项目期间，PC支持四个和 为47份出版物作出了贡献，比2006-2010年供资期间的15份出版物有所增加。在大多数情况下 最近一个财政年度，PC支持了26个成员，其中90%由同行审查资助的成员使用。 利用CCSG和机构基金提供的财政支持，筹委会继续提供最新情况- ART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.