PROTEIN IDENTIFICATION AND LOCALIZATION CORE

A.定义 蛋白质识别和定位(PIL)核心是一个重组的核心，它从 之前的两个核心，细胞生物学和细胞成像核心，专注于细胞 影像与多肽和蛋白质组学核心，具有提供合成的双重服务 多肽和质谱学鉴定蛋白质。虽然细胞生物学方法现在是 大多数实验室的标准和多肽合成都可以从大学和 商业消息来源，前两个核心的独特之处在于它们在识别和 定位蛋白质。新的PIL核心的结构保留和加强了这些方面 并建立在威廉姆斯博士、安德鲁斯博士和安永之间长期合作互动的基础上。 他们还基于这样一个事实，即这种类型的研究需要复杂而昂贵的设备，而不是 在单独的实验室中提供。PIL核心旨在提供有关 蛋白质通过质谱学，或者作为在研究人员实验室中纯化的单个蛋白质和 通常通过凝胶电泳法分离为条带或斑点，或全局分离为蛋白质的混合物 蛋白质复合体或细胞器。还提供了有关翻译后修改的信息 蛋白质和蛋白质含量的数量变化。一旦确定了一种蛋白质，核心可以 提供有关蛋白质在活细胞和固定细胞中的定位及其随时间变化的信息 以及对生理和病理生理扰动的响应。它以微观为中心 主要通过激光扫描共聚焦获得的数字信息的成像和定量分析 显微镜(LSCM)和多波长荧光成像，尽管传统的电子 固定和嵌入样品的显微镜也可用于精细结构分析。 现代生物医学研究既具有跨学科性质，又具有依赖性。 关于日益复杂的仪器和信息学。虽然目标是观察一种纸巾， 疾病，或有机体作为一个复杂的综合系统，我们仍在发展对 通常依赖于细胞类型的组件的结构和功能。通过映射到 随着基因组的发展，人们的注意力已经转移到更复杂的蛋白质组世界。完整的库存 大多数哺乳动物的蛋白质复合体或细胞器尚未完成，它们经常在 细胞类型、细胞周期和发育的不同阶段。从而识别出 蛋白质及其翻译后修饰、动态相互作用和细胞定位 和空间，并以定量的方式，是一项艰巨的任务，几乎所有的实验室科学家在美国 多肽中心在他们的研究中遇到了。这些问题可以通过以下工具解决 PIL核心，经常与分子生物学核心和国家基因组资源相一致 比如NCBI。 新的PIL核心建立在主任实验室的专业知识基础上，以及 现有的三个设施中的精密仪器，密歇根蛋白质组核心，形态学 和密歇根大学细胞与发育生物学系和成像实验室 糖尿病研究和培训中心(MDRTC)，该中心维护以下详细讨论的设施 精密的质谱学和高分辨率显微镜。这项安排是由 事实上，安德鲁斯博士领导着密歇根蛋白质组核心，威廉姆斯博士领导着显微镜和 MDRTC的图像分析实验室和Ernst博士与MIL有着长期的联系。全 三个实体已经建立了人员和充值结构，目前正由GI使用 中心成员。 而蛋白质识别和定位核心将利用现有的高度复杂的 仪器，它也是建立在超过90年的蛋白质组，细胞生物学和成像 威廉姆斯、安德鲁斯和安永博士的专业知识，主要致力于胃肠道组织。 包括Walker博士、Strahler博士和Nelson先生在内的经验丰富的核心员工将直接协助中心 调查人员和执行核心的程序。所有核心人员将进行示范和授课 向GI中心研究人员、实习生和技术人员提供技术培训。最后，核心的另一个使命是 发起、实施和传播蛋白质组学和细胞成像领域的新技术和创新技术。

A. DEFINITION The Protein Identification and Localization (PIL) Core is a restructured core that has evolved from aspects of two previous cores, the Cell Biology and Cell Imaging Core which focused on cellular imaging and the Peptides and Proteomics Core which had a dual service of providing synthetic peptides and identifying proteins by mass spectrometry. While cell biological methods are now standard in most laboratories and peptide synthesis can be obtained both from University and commercial sources, what was unique about the two previous cores was their synergy in identifying and localizing proteins. These aspects are preserved and enhanced by the structure of the new PIL Core and build on the longstanding collaborative interactions between Drs. Williams, Andrews and Ernst. They also build on the fact that this type of research requires complex and expensive equipment not available in individual laboratories. The PIL Core is designed to provide information on the identity of proteins by mass spectrometry, either as individual proteins purified in an investigator's laboratory and usually separated as a band or spot by gel electrophoresis or globally as a mixture of proteins in a protein complex or organelle. Information is also be provided on post-translational modifications of proteins and on quantitative changes in protein content. Once a protein is identified, the Core can provide information on the localization of the protein in live and fixed cells and their changes over time and in response to physiological and pathophysiological perturbation. It is centered around microscopic imaging and quantitative analysis of digital information obtained primarily by laser scanning confocal microscopy (LSCM) and multiwavelength fluorescence imaging, although traditional electron microscopy of fixed and embedded specimens is also available for fine structure analysis. Modern biomedical research is characterized both by its interdisciplinary nature and by its dependence on increasingly sophisticated instrumentation and informatics. Although the goal is to look at a tissue, disease, or organism as a complex integrated system, we are still developing an understanding of the structure and function of the components which are often cell type dependent. With the mapping of the genome, attention has shifted to the more complex world of the proteome. Complete inventories of most mammalian protein complexes or organelles are yet to be completed and they often vary between cell types, during the cell cycle, and during various stages of development. Thus identification of proteins, their post-translational modifications, dynamic interactions and cellular localization, all in time and space and in a quantitative manner, is a daunting task that almost all laboratory scientists in the GI Peptide Center encounter in their research. These questions can be approached through the tools of the PIL Core, very often in concert with the Molecular Biology Core and national genomic resources such as the NCBI. The new PIL Core is built around expertise in the director's laboratories, and the availability of sophisticated instrumentation in three existing facilities, the Michigan Proteome Core, the Morphology and Imaging Laboratory (MIL) of the Department of Cell and Developmental Biology and the Michigan Diabetes Research and Training Center (MDRTC) which maintain facilities discussed in detail below for sophisticated mass spectrometry and high resolution microscopy. This arrangement is facilitated by the fact that Dr Andrews directs the Michigan Proteome Core, Dr Williams directs the Microscopy and Image Analysis Laboratory of the MDRTC and Dr Ernst has a long standing affiliation with the MIL. All three entities have established personnel and a recharge structure and are currently being use by GI Center members. While the Protein Identification and Localization Core will make use of the available highly sophisticated instrumentation, it also is built around the over 90 years of proteomic, cell biology and imaging expertise of Drs. Williams, Andrews and Ernst which has been primarily devoted to GI Tissues. Experienced Core staff, including Drs. Walker, Strahler and Mr. Nelson, will directly assist Center Investigators and carry out the procedures of the Core. All Core personnel will demonstrate and teach techniques to GI Center investigators, trainees and technicians. Finally another mission of the Core is to initiate, implement and disseminate new and innovative techniques in proteomics and cell imaging.