Proteomics and Molecular Characterization

蛋白质组学共享资源 蛋白质组学和分子鉴定共享资源的总体目标是提供广泛的 为KCC调查人员提供一系列最先进的蛋白质分析服务。这些服务被分组为 三个主要领域：1)蛋白质组学和质谱学执行旨在帮助 研究人员在全球范围内识别和表征蛋白质(例如，表达水平，蛋白质复合体， 翻译后修改)。蛋白质鉴定用质谱仪(SELDI和 MALDI TOP和LC ESI)。目前，二维页面分析是目前使用的主要平台 确定实验样本之间的蛋白质表达差异，但我们已经做出了重大努力 将服务扩展到包括基于多维层析的方法。据我们估计， 这将是许多研究人员的首选工作流程，尤其是那些参与临床研究的人员。 因为高效液相分析所需的起始物质显著减少，而且可变性较小 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 正在为KCC成员开发胰蛋白酶多肽的iTRAQ)和18O标记。2)蛋白质和 多肽化学以非常合理的成本为KCC研究人员提供多肽合成服务。在……里面 除了常规肽之外，该资源还成功地合成了许多专业肽 资源已经成功地合成了极长的多肽(最多69个残基)以及环状 多肽。3)分子相互作用提供了表征蛋白质/蛋白质的仪器 互动。这些仪器包括Biacore 3000 SPR生物传感器、Kinexa 3000和等温 滴定量热计。除了这些部门所包含的服务外，共享资源 为用户提供访问SELDI-TOF质谱仪、MALDI-TOF质谱仪、台风 9400凝胶成像仪、紫外光谱仪以及超声波、真空离心机和 冷冻干燥。 这一共享资源提供的服务和设备在当前的 以及KCC调查人员未来的研究计划。这一共享资源经历了大量的管理 2006年12月的变化，通过整合两个活跃的研究实验室来改善资源，其 研究的重点是蛋白质组学和质谱学。我们的目标有三个：1)增加 为研究人员提供的服务数量；2)降低成本和增加按存储容量使用计费；3)带来 对资源进行新的检测。此外，随着资源的开始，使用率预计会增加 提供基于LC/MS蛋白质组工作流程和蛋白质微阵列的新服务。这些变化将 提高能力，同时降低运行该设施的总体成本。

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.