Core--Skin Proteomics

描述(申请人提供)：该核心的作用是提供蛋白质组学和质量 为UAB皮肤病研究中心的研究人员提供光谱支持。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，加上对组成各种项目产生的生物样品的蛋白质的质谱学鉴定。UAB综合癌症中心质谱学和蛋白质组学共享设施的现有仪器将用于皮肤蛋白质组学核心。由于这一资源得到了大量利用，发改委的资金将使调查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。核心将在实验设计和样本生成方面与国家发改委各种项目的调查人员合作，以确保最佳分析。因此，这个核心的具体目标是(1)优化细胞和组织样本中的蛋白质回收，(2)高分辨率蛋白质分离，(3) (4)通过软件(目前为PDQuest[BioRad])进行定性和定量的凝胶图像分析，以确定蛋白质表达的差异和/或翻译后修饰中的变化；(5)MALDI-TOF质谱学鉴定多肽，基于带有胰酶质量指纹图谱的数据库搜索；(6)NanoLC-电喷雾电离质谱学，用于多肽序列分析和共价修饰分析；(4)在蛋白质组学的实验设计和应用中进行培训，以及(7)蛋白质数据库和生物信息学的应用。总体而言，蛋白质组学核心的目标是(1)结合MALDI-TOF和其他质谱分析提供高分辨率的二维电泳和/或层析蛋白质分离，以及(2)识别多肽表达和/或多肽的差异 这可能是该中心调查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.