Skin Proteomics

描述（由申请人提供）：该核心的作用是提供蛋白质组学和质量 UAB皮肤病研究中心的研究人员的光谱支持。这将主要是以高分辨率分离复杂生物样品中的蛋白质的形式进行，同时用质谱法鉴定各种项目产生的生物样品中的蛋白质。UAB综合癌症中心质谱和蛋白质组学共享设施中的现有仪器将用于皮肤蛋白质组学核心。由于这一资源被大量利用，来自SDRC的资金将使研究人员能够为他们的皮肤相关项目获得技术帮助和专业知识。核心将与SDRC研究人员在实验设计和样品生成方面的各个项目合作，以确保最佳分析。因此，该核心的具体目标是（1）优化从细胞和组织样本中回收蛋白质，（2）高分辨率蛋白质分离，（3） 等电聚焦/SDS-PAGE电泳（3种不同的二维凝胶尺寸），（4）通过软件进行定性和定量凝胶图像分析（目前为PDQuest [BioRad]）以鉴定差异蛋白表达和/或翻译后修饰的改变，（5）基于用胰蛋白酶质量指纹的数据库搜索的多肽的MALDI-TOF质谱鉴定，（6）用于肽序列分析和共价修饰分析的NanoLC-电喷雾离子化质谱（NanoLC-Electrospray ionization mass spectrometry）蛋白质组学实验设计和应用的培训，以及（7）蛋白质数据库和生物信息学的使用。总的来说，蛋白质组学核心的目标是（1）提供与MALDI-TOF和其他质谱分析相结合的高分辨率二维电泳和/或色谱蛋白质分离，以及（2）鉴定多肽表达和/或蛋白质表达的差异。 这些修改可能是中心研究人员正在研究的各种皮肤病的基础。

DESCRIPTION (provided by applicant): The role of this Core is to provide proteomics and mass spectrometry support for the investigators in the UAB Skin Diseases Research Center. This will be principally in the form of high-resolution separation of proteins in complex biological samples, coupled with mass spectrometry identification of proteins that comprise the biological samples generated by the various projects. Existing instrumentation m the UAB Comprehensive Cancer Center Mass Spectrometry and Proteomics Shared Facility will be used for the Skin Proteomics Core. Because this resource is heavily utilized, funding from the SDRC will enable investigators to obtain the technical help and expertise for their skin related projects. The Core will work with the SDRC investigators of the various projects in experimental design and sample generation, to ensure optimal analysis. Thus, the specific aims of this Core are (1) Optimization of protein recovery from cells and tissue specimens, (2) High resolution protein separation, (3) Isoelectric focusing/SDS-PAGE electrophoresis (on three different 2 nd dimension gel sizes), (4) Qualitative and quantitative gel image analysis via software (currently PDQuest [BioRad]) to identify differential protein expression and/or alterations in posttranslational modifications, (5) MALDI-TOF mass spectrometric identification of polypeptides based on database searching with tryptic mass fingerprints, (6) NanoLC-electrospray ionization mass spectrometry for peptide sequence analysis and analysis of covalent modifications Training in the experimental design and application of proteomics, and (7) use of protein databases and bioinformatics. Overall, the goals of the proteomics core are (1) to provide high-resolution two dimensional electrophoretic and/or chromatographic protein separations coupled with MALDI-TOF and other mass spectrometry analyses, and (2) to identify differences in polypeptide expression and/or modifications that may be the basis for various dermatological conditions being studied by the Center investigators.