Proteomics Shared Resource

Proteomics共享资源项目摘要 马约诊所癌症中心（MCCC）蛋白质组学共享资源（PRO）是一个提供全方位服务的机构， 提供肽合成、蛋白质分析、蛋白质和肽分离、质谱分析 光谱学、蛋白质组学应用以及项目规划、样品制备、数据采集方面的专业知识 和分析PRO占地4920平方米。英尺在医学科学大楼的三楼， 位于罗切斯特（MCR）研究中心的马约诊所。8名专业干事知识渊博， 提供服务的专业知识。专业主任，丹尼尔J麦考密克博士，提供有远见的方向， 有关所有运营、人员配备和服务的规划、招聘和执行决策。使用 机构委员会和中心超过33年，麦考密克博士已经能够提供科学的 指导和支持高端核心仪器，以及蛋白质化学，蛋白质组学的新技术 和质谱（MS），以支持MCCC成员在癌症研究中的计划。上一 在供资期间，6个新的服务项目（例如，无标记差异蛋白质组学，扩展磷酸化蛋白质组学， SCX分级分离和TiO 2富集，FFPE和冷冻组织的蛋白质组学分析，高/低pH LC 样品分级分离、从头MS/MS蛋白质测序和复杂聚糖的MS分析）。 开发旨在支持MCCC成员的蛋白质分析需求。科学成就包括： 在骨髓癌中鉴定与TAT 2蛋白相互作用的关键蛋白;开发 在PDX细胞系中抑制TGF β 1作用（通过SMAD信号传导）并防止胶质母细胞瘤迁移; 关键磷蛋白（例如，CXCR 4）在Alisertib治疗的人乳腺癌中下调; 肺“干性”表型转录激活因子SOX 2磷酸化位点定位 鳞状细胞癌;用于过继免疫中体外激活幼稚T细胞的癌蛋白抗原的合成 治疗癌症的T细胞疗法（例如，胰腺）;和差异分析的蛋白质激活SPINK 1在 卵巢癌细胞增殖MCCC成员的PRO使用率一直很高，总平均使用率为 51%由99名成员（72名同行评审）在当前资助期间（2013-2017年）。PRO由MCCC使用 来自所有3个研究中心的成员，包括佛罗里达州马约诊所的12名成员和马约诊所的5名成员 在亚利桑那州专业档案室为MCCC成员提供免费咨询、项目设计和数据解释， ，并以收费服务方式提供服务;项目和服务要求的标准表格载于 资源的内部网站。专业干事的许多服务与外部核心服务的费率比较， 供应商，于2017年完成，并表示核心费率极具竞争力。的未来方向 PRO包括：使用先进的MS仪器和数据独立的深度测序蛋白质组学 采集（DIA）方法;自顶向下MS蛋白质测序;离子迁移率分离（IMS）质谱; 基因组学/蛋白质组学数据集的蛋白质组学整合，以及糖蛋白的MS分析。

PROTEOMICS SHARED RESOURCE PROJECT SUMMARY The Mayo Clinic Cancer Center (MCCC) Proteomics Shared Resource (PRO) is a full-service facility that provides essential services in peptide synthesis, protein analysis, protein and peptide separation, mass spectrometry, proteomics applications, and expertise in project planning, sample preparation, data acquisition and analysis. The PRO occupies a total of 4920 sq. ft. on the 3rd floor of the Medical Sciences Building, and is located at the Mayo Clinic in Rochester (MCR) site. The 8 PRO staff members have broad knowledge and expertise in the services provided. The PRO Director, Dr. Daniel J. McCormick, provides visionary direction, planning, recruitment, and executive decisions regarding all operations, staffing, and services. Working with institutional committees and centers for over 33 years, Dr. McCormick has been able to provide scientific direction and support for high-end core instrumentation, and new technologies in protein chemistry, proteomics and mass spectrometry (MS) to support Programs of MCCC members in cancer research. In the previous funded period, 6 new service-lines (e.g., label-free differential proteomics, expanded phosphoproteomics by SCX fractionation and TiO2 enrichment, proteomics analysis of FFPE and Frozen tissue, high/low pH LC sample fractionation, de novo MS/MS protein sequencing, and MS analysis of complex glycans) were developed to support the protein analysis needs of MCCC members. Scientific achievements include: identification of key proteins that interact with TAT2 protein in myeloid cancers; development of agents that inhibit TGFß action (via SMAD signaling) and prevent glioblastoma migration in PDX cell lines; identification of key phosphoproteins (e.g., CXCR4) downregulated in Alisertib-treated human breast adenocarcinoma; mapping sites of phosphorylation in SOX2, a transcriptional activator of “stemness” phenotype in lung squamous cell carcinoma; synthesis of oncoprotein antigens for in vitro activation of naïve T-cells in adoptive T-cell therapies to treat cancer (e.g., pancreatic); and differential analysis of proteins activated by SPINK1 in ovarian cancer proliferation. PRO utilization by MCCC members has been strong with a total average usage of 51% by 99 members (72 peer-reviewed) in the current funded period (2013-2017). The PRO is used by MCCC members from all 3 sites, including 12 members from Mayo Clinic in Florida and 5 members from Mayo Clinic in Arizona. The PRO provides free consultation, project design, and data interpretation to MCCC members, and provides its services on fee-for-service basis; standardized forms for project and service requests are on the resources' internal web site. Comparison of rates for many services of the PRO to external cores and vendors, was completed in 2017 and indicated that core rates were highly competitive. Future directions of the PRO include: deep sequencing proteomics with advanced MS instrumentation and data-independent acquisition (DIA) methods; top-down MS protein sequencing; ion mobility separation (IMS) mass spectrometry; proteogenomics integration of genomics/proteomics data sets, and MS analysis of glycoproteins.