Proteomics Core

该核心将为所有项目提供“最先进的”蛋白质组学和质谱分析支持（J. Zheng， H. Gendelman和Y. Persidsky）在这份提案中。这将包括神经元祖细胞的蛋白质谱 细胞、单核细胞衍生的巨噬细胞、小胶质细胞以及与影响 病毒和细胞蛋白质的变化，脑单核细胞炎症反应和发育 神经艾滋病的治疗我们将使用差异凝胶双向电泳（2DE） 电泳（DICE）（2DE DICE）作为主要分析方法。为了补充这些既定的 方法，我们将使用ABI 4800 MALDI-TOF/TOF（Applied Biosystems，Inc.）用于快速精确的质量 分析.翻译后修饰如磷酸化将用QQ-LIT（QTrap 4000， Applied Biosystems，Inc.）。此外，我们还有定量质谱分析系统， 同位素编码亲和标签（ICAT）、相对和绝对定量同位素标签（iTRAQ）和 用于蛋白质标记或标签的细胞培养物中氨基酸稳定同位素标记（SILAC）。我们正在进行的 与Beckman-Coulter，Inc.合作通过获得一种 Proteomel_ab <$PF 2D蛋白质分级系统用于分析复杂蛋白质样品。采取 总之，这个核心是从简单的表面增强激光解吸电离分析发展而来的， 最初的计划项目建议书，成为一个大学范围内的良好集成的核心设施，这有助于 极大地促进了申请中包含的所有拟议研究。

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.