CHARACTERIZATION OF GLOBAL YEAST QUANTITATIVE PROTEOME DATA GENERATED FROM THE W

这个子项目是许多利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 复杂蛋白质混合物的定量蛋白质组学分析是一种新兴的技术， 具有挑战性但可行的研究细胞功能的系统级方法。本研究 提出了一个大规模的比较分析蛋白质丰度从酵母蛋白裂解物 来源于野生型酵母和缺乏Snf1关键组分的酵母菌株 激酶复合物四种不同的菌株在良好控制的恒化器下生长 条件多维蛋白质鉴定技术，随后使用 无论是光谱计数或稳定同位素标记的方法来确定相对 菌株之间蛋白质表达水平的变化。共检测到2388种蛋白质， 相对定量，超过350种蛋白质被发现有显着不同， 当使用稳定同位素时，两种比较菌株之间的表达水平 标签策略基于稳定同位素标记的定量方法被发现是 当复杂的蛋白质混合物含有 分析小的表达变化。其中稳定同位素 标记和光谱计数，发现差异背后的主要原因是 缺乏对具有低光谱计数的蛋白质的可重复采样。功能 分类的相对蛋白质表达差异，发生在Snf1缺陷 菌株揭示了由这种重要的细胞调节的广泛的生物过程。 激酶。

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.