Optimized protein extraction for quantitative proteomics of yeasts.
Optimized protein extraction for quantitative proteomics of yeasts.
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优化的蛋白质提取酵母定量蛋白质组学。
DOI：
10.1371/journal.pone.0001078
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发表时间：
2007-10-24
期刊：
影响因子：
3.7
通讯作者：
von der Haar, Tobias
中科院分区：
文献类型：
--
作者：
von der Haar, Tobias
细胞内蛋白水平的绝对定量在技术上要求很高，但最近变得更加突出，因为系统生物学和代谢控制分析等新方法需要了解这些参数。目前从酵母细胞中提取蛋白质的方法可能会由于不完全提取或选择性提取而在定量过程中引入伪影。本研究建立了一种基于化学裂解和SDS和尿素同时溶解的酿酒酵母蛋白质提取新工艺，该工艺可提取绝大部分蛋白质，并达到明显的完整程度。该方法可用于不同的酵母菌种类和不同的生长条件，适用于96孔格式的高通量提取，所得提取物可以很容易地后处理用于非sds兼容的程序，如2D凝胶电泳。已经开发了一种改进的定量蛋白质提取方法，该方法可以在定量蛋白质组学实验中去除一些人工制品的来源，同时允许新的应用类型。
The absolute quantification of intracellular protein levels is technically demanding, but has recently become more prominent because novel approaches like systems biology and metabolic control analysis require knowledge of these parameters. Current protocols for the extraction of proteins from yeast cells are likely to introduce artifacts into quantification procedures because of incomplete or selective extraction. We have developed a novel procedure for protein extraction from S. cerevisiae based on chemical lysis and simultaneous solubilization in SDS and urea, which can extract the great majority of proteins to apparent completeness. The procedure can be used for different Saccharomycetes yeast species and varying growth conditions, is suitable for high-throughput extraction in a 96-well format, and the resulting extracts can easily be post-processed for use in non-SDS compatible procedures like 2D gel electrophoresis. An improved method for quantitative protein extraction has been developed that removes some of the sources of artefacts in quantitative proteomics experiments, while at the same time allowing novel types of applications.
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影响因子：
2.9
作者：
Remacha, M;JimenezDiaz, A;Ballesta, JPG
通讯作者：
Ballesta, JPG
DOI：
10.1073/pnas.76.9.4350
发表时间：
1979-01-01
影响因子：
11.1
作者：
TOWBIN, H;STAEHELIN, T;GORDON, J
通讯作者：
GORDON, J
影响因子：
56.9
作者：
CHERNOFF, YO;LINDQUIST, SL;LIEBMAN, SW
通讯作者：
LIEBMAN, SW
影响因子：
64.8
作者：
Newman, John R. S.;Ghaemmaghami, Sina;Weissman, Jonathan S.
通讯作者：
Weissman, Jonathan S.
影响因子：
64.8
作者：
LAEMMLI, UK
通讯作者：
LAEMMLI, UK