Fourier transform-ion cyclotron-resonance mass spec

描述(由申请人提供)：目前的提案将通过提供资金购买傅立叶变换离子回旋共振质谱仪(FTMS)来帮助扩大系统生物学研究所(ISB)的蛋白质组学设施。目前，蛋白质组学设施的重点是大规模应用一种新的方法，对细胞和组织中存在的蛋白质进行系统鉴定和准确定量，从而检测生物过程或医疗条件所特有的全球蛋白质表达谱的动态变化。该中心和NIH支持的生物医学研究人员之间存在着大量的合作，在这项提案中概述了各种领域，如丙型肝炎的干扰素耐药性，前列腺癌中的雄激素依赖，干细胞生物学，识别与Tollike受体相关的蛋白质，Myc癌蛋白的功能分析，酵母中基因表达的细胞周期调控，胸腺细胞生物学，以及与T-Ceil调节蛋白LAT相关的蛋白质分析和相关的磷酸化事件。除了NIH支持的这些项目外，该设施与ISB内外的合作者之间正在进行另外12项合作。该设施中的FTMS仪器的可用性将允许对每个项目中的单个蛋白质及其翻译后修饰(PTM)和整个蛋白质组进行更全面的表征，而不是使用大西雅图地区现有的任何其他类型的质谱仪。FTMS仪器无损离子检测的独特功能在三个关键的质谱学参数上提供了优异的性能，这将有助于我们的生物医学研究：1)在1000u处0.1-1.0ppm的质量精度允许更高的置信度，例如，通过质量指纹方法识别多肽中的蛋白质，2)&GT的质量分辨率；100000(50%mIDm)在1000u时提供了在复杂混合物表征领域的非凡能力，其中在原油混合物中检测到多达3000个组分(参考马歇尔)，并且3)信噪比增强允许在低阿托摩尔区域(马歇尔)中检测和碎裂多肽。一台FTMS仪器在这三个领域的数据都远远超过了目前可用的任何其他质谱仪的相同能力。此外，在大西雅图地区没有FTMS仪器可供我们或我们的合作者使用。

DESCRIPTION (provided by applicant): The current proposal will help to expand the Institute for System Biology's (ISB) Proteomics facility by providing capital to purchase a Fourier transform-ion cyclotron-resonance mass spectrometer (FTMS). Currently, the Proteomics facility focus is on large scale application of a novel approach to the systematic identification and accurate quantitation of the proteins present in cells and tissues and therefore detection of dynamic changes in global protein expression profiles that are characteristic for biological processes or medical conditions. Numerous collaborations exist between the facility and NIH supported biomedical researchers that are outlined in this proposal covering areas as diverse as interferon resistance to hepatitis C, androgen dependence in prostate cancer, stem cell biology, identification of proteins associated with Tollike receptors, functional analysis of the Myc oncoprotein, cell-cycle regulation of gene expression in the yeast Saccharomyces cerevisiae, thymocyte biology, and analysis of the proteins associated with the T-ceIl regulatory protein LAT and associated phosphorylation events. In addition to these NIH supported projects another 12 collaborations are ongoing between the facility with collaborators inside and out of the ISB. The availability of FTMS instrumentation in the facility will allow more thorough characterization of individual proteins and their post-translational modifications (PTM) and whole proteomes for each of these projects than is possible with any of the other types of mass spectrometers present in the greater Seattle area. The unique features of non-destructive ion detection by FTMS instrumentation provide exceptional performance in three critical mass spectrometric parameters that will benefit our biomedical research: 1) mass accuracy of 0.1 -1.0 ppm at 1000 u allow higher confidence in identifications of proteins from peptides by, for instance, the mass fingerprint approach, 2) mass resolution of >1 00,000 (50% mIDm) at 1000 u provides extraordinary capability in the area of complex mixture characterization where as many as 3000 components have been detected in a crude oil mixture (REF Marshall) and 3) signal-to-noise enhancement allows detection and fragmentation of peptides in the low attomole regime (Marshall). Data from an FTMS instrument in each of these three areas far exceeds the same capability from any other mass spectrometer that is currently available. Furthermore there are no FTMS instruments in the greater Seattle area for us, or our collaborators, to use.