Proteomics Core

该核心将为所有项目提供“最先进的”蛋白质组和质谱学支持(J·郑， H.Gendelman和Y.Persidsky)在这份提案中。这将包括神经元前体细胞的蛋白质图谱。 细胞、单核细胞来源的巨噬细胞、小胶质细胞以及细胞与细胞之间的相互作用 脑单核细胞炎症反应和发育过程中病毒和细胞蛋白的变化 神经艾滋病的治疗学。我们将使用差异凝胶的双向凝胶(2DE) 电泳(骰子)(2DE骰子)作为主要的仿形方法。为了补充这些已建立的 方法：我们将使用ABI4800 MALDI-TOF/TOF(应用生物系统公司)对于快速和精确的质量 分析。翻译后修饰如磷酸化将使用QQ-LIT(QTRap4000， 应用生物系统公司)。此外，我们还有定量质谱学系统，结合了 同位素编码亲和标签(ICAT)、相对和绝对量化的同位素标签(ITRAQ)？以及 细胞培养中氨基酸稳定同位素标记(SILAC)用于蛋白质标记或标记。我们正在进行的 与Beckman-Coulter，Inc.的合作通过收购一家 Proteomel_ab？PF2D蛋白质分级系统用于分析复杂蛋白质样品。已被占用 总而言之，这个核心是从包括在 最初的方案项目建议成为全大学范围内综合良好的核心设施，这将有助于 极大地促进了申请中包括的所有拟议研究。

This core will provide "state of the art" proteomic and mass spectrometry support for all projects (J. Zheng, H. Gendelman, and Y. Persidsky) in this proposal. This will include protein profiling for neuronal progenitor cells, monocyte-derived macrophages, microglia as well as cell-cell interactions relevant to studies that affect virus and cell-protein changes in brain mononuclear cell inflammatory responses and for developmental therapeutics in NeuroAIDS. We will use 2-dimensional electrophoresis (2DE) with Difference Gel Electrophoresis (DICE) (2DE DICE) as a primary profiling approach. To complement these established methods, we will use an ABI4800 MALDI-TOF/TOF (Applied Biosystems, Inc.) for rapid and precise mass analysis. Post-translational modifications such as phospnorylation will be done with a QQ-LIT (QTrap4000, Applied Biosystems, Inc.). In addition, we have quantitative mass spectrometry systems that incorporates isotope-coded affinity tags (ICAT), isotope Tags for Relative and Absolute Quantification (iTRAQ)¿, and Stable Isotope-labeling with Amino Acids in Cell culture (SILAC) for protein labeling or tagging. Our ongoing collaborations with Beckman-Coulter, Inc. have extended our protein capabilities with the acquisition of a Proteomel_ab¿PF2D Protein Fractionation System for the profiling complex protein samples. Taken together, this core has evolved from simple surface enhanced laser desorption ionization assays included in the initial program project proposal to become a university-wide well-integrated core facility, which serves to greatly facilitate all the proposed research included in the application.