Core 012 (797) Proteomics Core

核心 012 (797) 蛋白质组学核心

基本信息

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
项目类别：
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

项目摘要

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.

项目概要蛋白质组学核心 (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分析技术来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) 核心于 2003 年建立。并在2011年评审中被评为“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流体。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。蛋白质组学核心的总体目标是：1）与成员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； 3) 培训成员及其工作人员 PC 由四名全职工作人员组成，并提供液体。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 (LC-MRM) 用于目标定量。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）、药物靶点（化学蛋白质组学）、信号传导（磷酸化蛋白质组学）、蛋白质组学广泛的乙酰化或泛素化、PTM 的连续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 敲低、CRISPR 敲除）。 PC 使用代谢标记 (SILAC) 或化学标记 (iTRAQ/TMT) 提供样本多重分析最后，PC 使用发现蛋白质组学的集成流程来进行比较蛋白质组学技术。探索癌症生物学和目标定量，以实现高精度的生物标志物测量从模型系统转化为 PC 与分子基因组学核心的交互。 (MGC) 对于联合蛋白质组学分析是必要的；并且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的下游相互作用。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 支持四个和多个用户。资助了 47 种出版物，比 2006 年至 2010 年资助期间的 15 种出版物有所增加。最近一个财年，PC 支持了 26 个成员，其中 90% 的使用量由同行评审资助的成员使用。利用 CCSG 和机构资金提供的财政支持，PC 继续提供最先进的服务先进的设备、技术和服务，以满足MCC会员当前和未来的需求。