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PROTEIN IDENTIFICATION AND LOCALIZATION CORE

蛋白质鉴定和定位核心

基本信息

批准号：
8385555
负责人：
JOHN A WILLIAMS
金额：
$ 13.42万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/8385555
关键词：
3-Dimensional Actins Affect Affinity Algorithms Amylases Animals Antibodies Area Attention Biological Biomedical Research Blood capillaries Cancer Center Cell Cycle Cell Fraction Cell Line Cell Nucleus Cells Cellular biology Chemicals Chemistry Citrates Clinical Investigator Collection Color Complex Computer software Computers Computing Methodologies Confocal Microscopy Consultations Coomassie blue Copper Core Protein Coupled Cryoultramicrotomy Cybernetics Data Data Analyses Data Files Databases Dependence Development Developmental Cell Biology Devices Diabetes Mellitus Dimensions Disease Dyes Educational process of instructing Electron Microscopy Endocytosis Energy Transfer Enzymes Equipment Equipment and supply inventories Evaluation F-Actin Fees Fixatives Fluorescence Microscopy Fluorescence Resonance Energy Transfer Fluorescent Antibody Technique Fluorescent Probes Frequencies Frozen Sections Fura-2 Gel Genome Mappings Genomics Goals Gold Hand High Pressure Liquid Chromatography Hormones Hour Housing Human Resources Image Image Analysis Imagery Immunofluorescence Immunologic Immunohistochemistry Individual Informatics Instruction Isotopes Label Laboratories Laboratory Research Laboratory Scientists Laser Scanning Confocal Microscopy Lasers Lead Life Light Location Mammalian Cell Manuals Maps Mass Spectrum Analysis Measurement Measures Methodology Methods Michigan Microscope Microscopic Microscopy Mission Mitochondria Modeling Modification Molecular Molecular Biology Monitor Morphology Nature Nickel Nuclear Organelles Organism Paraffin Embedding Pathology Peptide Hydrolases Peptide Synthesis Peptides Perfusion Phase Phosphopeptides Phosphoproteins Phosphorylated Peptide Phosphorylation Phosphorylation Site Physiological Pilot Projects Plant Resins Post-Translational Modification Site Post-Translational Protein Processing Procedures Process Property Protein Chemistry Proteins Proteome Proteomics Protocols documentation Publications Publishing Recipe Research Research Design Research Personnel Research Training Resolution Resources Sampling Series Services Signal Pathway Silver Site Source Specimen Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Spottings Staging Staining method Stains Structure System Techniques Time Tissues Tolonium chloride Training Transmission Electron Microscopy Tube Universities Walkers base capillary cell fixation cell fixing cell type cellular imaging cost cryostat design digital digital imaging epon experience fluorescence imaging fluorescence microscope fluorophore gel electrophoresis immunoaffinity chromatography immunocytochemistry improved innovation instrument instrumentation interest mass spectrometer member novel operation pH gradient protein complex ratiometric reconstruction research facility research study response sample fixation scaffold screening small molecule sodium-binding benzofuran isophthalate spurr resin synthetic peptide tandem mass spectrometry tissue preparation tool uranyl acetate vector zirconium oxide

项目摘要

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.

答:定义