Core 012 (797) Proteomics Core

核心 012 (797) 蛋白质组学核心

• 批准号: 10230154
• 负责人: John M Koomen
• 金额: $ 0.45万
• 依托单位: H. LEE MOFFITT CANCER CTR & RES INST
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-02-18 至 2022-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10230154
• 关键词: ART protein; Acetylation; Address; Affinity Chromatography; Analytical Chemistry; Animal Model; Antibodies; Applications Grants; Biological Assay; Biological Markers; Biological Models; Biological Process; Biostatistics Core; Budgets; Cancer Biology; Cancer Center; Cancer Center Support Grant; Cell Line; Chemicals; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Complement; Complex; Consult; Data; Data Analyses; Development; Doctor of Philosophy; Drug Targeting; Education; Environment; Equipment; Evaluation; Event; Experimental Designs; Financial Support; Florida; Formalin; Fractionation; Freezing; Funding; Future; Genomics; Genotype; Goals; Group Meetings; Immunoprecipitation; In Situ; Individual; Informatics; Institution; Instruction; Knock-out; Knowledge; Label; Laboratories; Letters; Link; Liquid Chromatography; Maintenance; Malignant Neoplasms; Manuscripts; Mass Spectrum Analysis; Measurement; Measures; Metabolic; Methods; Microscopy; Molecular; Mutation; Patients; Peer Review; Peptide Synthesis; Peptides; Persons; Pharmaceutical Preparations; Pharmacotherapy; Phenotype; Post-Translational Protein Processing; Protein Chemistry; Proteins; Proteome; Proteomics; Publications; Reporting; Research; Research Project Grants; Resource Sharing; Sampling; Services; Signal Transduction; Small Interfering RNA; Specimen; Techniques; Technology; Testing; Time; Tissues; Training; Training Programs; Translating; Treatment Protocols; Tumor Biology; Tumor Tissue; Ubiquitination; Universities; base; biomarker development; cancer biomarkers; cancer cell; cohort; community center; comparative; data mining; design; experience; experimental study; innovation; instrumentation; knock-down; laboratory experience; meetings; member; metabolomics; multiple reaction monitoring; neoplastic cell; phosphoproteomics; programs; protein complex; protein expression; protein protein interaction; proteogenomics; recruit; response; tandem mass spectrometry; treatment strategy; tumor

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.

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，用于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（PTMS）。核心成立于2003年 并在2011年的评论中获得了“杰出”的评分。可以用细胞系和动物进行实验 模型以及患者标本，包括潜水组织类型和生物流体。这些范围 研究范围从单个蛋白质的详细分子表征到整个蛋白质组的分析。这 蛋白质组学核心的总体目的是：1）与设计成员咨询和合作 执行蛋白质组学实验； 2）提供仪器和训练有素的员工来支持成员； 3）培训成员及其员工。 PC已完成四名全职员工，并提供液体 色谱串联质谱（LC-MS/MS）肽测序，用于发现蛋白质组学，和 液相色谱 - 形成反应监测质谱法（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），信号传导（磷蛋白质组学），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使用代谢标签（SILAC）或化学标签（ITRAQ/TMT）提供样品多路复用 比较蛋白质组学的技术。最后，PC使用Discovery蛋白质组学的集成管道进行 探索癌症生物学和目标数量，以进行高度精确的生物标志物测量值 从模型系统转换为患者标本。 PC与分子基因组核心的相互作用 （MGC）对于蛋白质分析组合是必需的；并与佛罗里达大学东南大学 综合代谢组学中心支持蛋白质代谢组学。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支持四个和四个的用户 贡献了47个出版物，比2006 - 2010年融资期的15个出版物增加。最多 最近的财政年度，PC支持了26名成员，同行评估资助的成员的总使用量的90％。 使用CCSG和机构基金提供的财务支持，PC继续提供最新的 艺术设备，技术和服务，以满足MCC成员当前和未来的需求。