CHARACTERIZATION OF GLOBAL YEAST QUANTITATIVE PROTEOME DATA GENERATED FROM THE W

W 生成的全球酵母定量蛋白质组数据的表征

• 批准号: 8171236
• 负责人: James Akira Wohlschlegel
• 金额: $ 0.95万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8171236
• 关键词: Biological; Biological Process; Cell physiology; Complex; Computer Retrieval of Information on Scientific Projects Database; Data; Funding; GLC2 protein; Grant; Institution; Phosphotransferases; Protein Analysis; Proteins; Proteome; Proteomics; Relative (related person); Research; Research Personnel; Resources; Sampling; Source; Stable Isotope Labeling; System; Technology; United States National Institutes of Health; Yeasts; base; comparative; protein complex; protein expression; yeast protein

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The quantitative proteomic analysis of complex protein mixtures is emerging as a technically challenging but viable systems-level approach for studying cellular function. This study presents a large-scale comparative analysis of protein abundances from yeast protein lysates derived from both wild-type yeast and yeast strains lacking key components of the Snf1 kinase complex. Four different strains were grown under well-controlled chemostat conditions. Multidimensional protein identification technology followed by quantitation using either spectral counting or stable isotope labeling approaches was used to identify relative changes in the protein expression levels between the strains. A total of 2388 proteins were relatively quantified, and more than 350 proteins were found to have significantly different expression levels between the two strains of comparison when using the stable isotope labeling strategy. The stable isotope labeling based quantitative approach was found to be highly reproducible among biological replicates when complex protein mixtures containing small expression changes were analyzed. Where poor correlation between stable isotope labeling and spectral counting was found, the major reason behind the discrepancy was the lack of reproducible sampling for proteins with low spectral counts. The functional categorization of the relative protein expression differences that occur in Snf1-deficient strains uncovers a wide range of biological processes regulated by this important cellular kinase.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 复杂蛋白质混合物的定量蛋白质组分析正在成为一种技术 用于研究细胞功能的具有挑战性但可行的系统级方法。这项研究 对酵母蛋白裂解物的蛋白质丰度进行了大规模比较分析 源自野生型酵母和缺乏 Snf1 关键成分的酵母菌株 激酶复合物。四种不同的菌株在良好控制的恒化器下生长 状况。多维蛋白质鉴定技术，然后使用 使用光谱计数或稳定同位素标记方法来识别相对 菌株之间蛋白质表达水平的变化。总共2388个蛋白质 相对定量，发现超过350种蛋白质具有显着差异 使用稳定同位素时比较两个菌株之间的表达水平 标签策略。发现基于稳定同位素标记的定量方法 当复杂的蛋白质混合物含有 分析了小的表达变化。稳定同位素之间的相关性较差 发现标签和光谱计数，差异背后的主要原因是 缺乏对低光谱计数蛋白质的可重复采样。功能性的 Snf1 缺陷中发生的相对蛋白质表达差异的分类 菌株揭示了受这种重要细胞调节的广泛生物过程 激酶。