Proteomics and Molecular Characterization

蛋白质组学共享资源 蛋白质组学和分子表征共享资源的总体目标是提供广泛的 为KCC研究人员提供一系列最先进的蛋白质分析服务。这些服务分为 三个主要领域：1）蛋白质组学和质谱分析进行旨在帮助 研究人员在全球范围内鉴定和表征蛋白质（例如表达水平，蛋白质复合物， 翻译后修饰）。蛋白质鉴定用质谱法（SELDI和SDS-PAGE）完成。 MALDI TOP和LC ESI）。目前，二维PAGE分析是用于分析蛋白质的主要平台。 定义实验样本之间蛋白质表达的差异，但我们已经做出了重大努力， 扩大服务范围，以包括基于多维色谱的方法。据我们估计， 这将是许多研究者的首选工作流程，尤其是那些参与临床研究的研究者 因为HPLC分析需要显著更少的起始材料并且因为更小的可变性 与基于凝胶的分离相比。用于同位素标记的商业试剂盒（例如SILAC、ICAT和 ITRAQ）以及18 O标记的胰蛋白酶肽正在开发的KCC成员。2)蛋白质和 肽化学以非常合理的成本为KCC研究人员提供肽合成服务。在 除了常规肽外，该资源还成功合成了许多特殊肽。 资源已成功地合成极长的肽（高达69个残基），以及环 缩氨酸3)分子相互作用提供了表征蛋白质/蛋白质的仪器 交互.这些仪器包括Biacore 3000 SPR生物传感器、Kinexa 3000和等温生物传感器。 滴定热量计。除了这些部门提供的服务外， 为用户提供SELDI-TOF质谱仪、MALDI-TOF质谱仪、Typhoon 9400凝胶成像仪、紫外光谱仪以及超声处理、真空离心和 冷冻干燥 这种共享资源提供的服务和设备在当前的环境中发挥着重要作用。 以及KCC调查人员的未来研究计划。这一共享资源经历了重要的管理 2006年12月，通过整合两个活跃的研究实验室来改善资源， 研究的重点是蛋白质组学和质谱法。我们的目标有三个方面：1）增加 研究人员可获得的服务数量; 2）降低成本并增加退款;以及3） 向资源添加新工具。此外，随着资源开始使用， 提供基于LC/MS蛋白质组学工作流程和蛋白质微阵列的新服务。这些变化将 提高能力，同时降低运行设施的总体成本。

PROTEOMICS SHARED RESOURCE The overall goal of the Proteomics and Molecular Characterization Shared Resource is to provide a wide range of state-of-the-art protein analysis services for KCC investigators. These services are grouped into three major areas: 1) Proteomics and Mass Spectrometry performs experiments designed to help investigators identify and characterize proteins on a global scale (e.g. expression levels, protein complexes, post-translational modifications). Protein identification is accomplished with mass spectrometry (SELDI and MALDI TOP and LC ESI). Currently, 2-Dimensional PAGE analysis is the predominant platform used to define differences in protein expression between experimental samples, but we have made significant efforts to expand the services to include multidimensional chromatography-based approaches. In our estimation, this will be the preferred workflow for many investigators especially those involved with clinical studies because the HPLC analysis requires significantly less starting material and because of less variability compared with the gel based separations. Commercial kits for isotopic labeling (e.g. SILAC, ICAT and ITRAQ) as well as 18O labeling of tryptic peptides are being developed for KCC members. 2) Protein and Peptide Chemistry provides peptide synthesis services for KCC investigators at very reasonable costs. In addition to routine peptides, the resource has successfully synthesized many specialized peptidesThe resource has been successful in synthesizing extremely long peptides (up to 69 residues), as well as cyclic peptides. 3) Molecular Interaction provides access to instrumentation for characterizing protein/protein interactions. These instruments include a Biacore 3000 SPR biosensor, Kinexa 3000, and an Isothermal Titration Calorimeter. In addition to the services contained within these Divisions, the shared resource provides users access to a SELDI-TOF mass spectrometer, a MALDI-TOF mass spectrometer, Typhoon 9400 Gel Imager, UV spectrometer, as well as equipment for sonication, vacuum centrifugation, and lyophilization. The services and equipment made available by this shared resource play an important role in the current and future research plans of KCC investigators. This shared resource underwent significant management changes in December 2006 to improve the resource by integrating two active research laboratories whose research is focused on proteomics and mass spectrometry. Our goals are threefold: 1) to increase the number of services available to researchers; 2) to reduce costs and increase chargebacks; and 3) to bring new instrumentation to the resource. Additionally, usage is expected to increase as the resource begins offering new services based on LC/MS proteomics workflows and protein microarrays. These changes will improve the capabilities while decreasing the overall costs of running the Facility.