Weckwerth Wolfram, Wenzel Kathrin, Fiehn Oliver
Max-Planck-Institut für Molekulare Pflanzenphysiologie, Golm, Germany.
Proteomics. 2004 Jan;4(1):78-83. doi: 10.1002/pmic.200200500.
A novel extraction protocol is described with which metabolites, proteins and RNA are sequentially extracted from the same sample, thereby providing a convenient procedure for the analysis of replicates as well as exploiting the inherent biological variation of independent samples for multivariate data analysis. A detection of 652 metabolites, 297 proteins and clear RNA bands in a single Arabidopsis thaliana leaf sample was validated by analysis with gas chromatography coupled to a time of flight mass spectrometer for metabolites, two-dimensional liquid chromatography coupled to mass spectrometry for proteins, and Northern blot analysis for RNA. A subset of the most abundant proteins and metabolites from replicate analysis of different Arabidopsis accessions was merged to form an integrative dataset allowing both classification of different genotypes and the unbiased analysis of the hierarchical organization of proteins and metabolites within a real biochemical network.
本文描述了一种新型提取方案，可从同一样品中依次提取代谢物、蛋白质和RNA，从而为重复分析提供了一种便捷的方法，同时利用独立样品的固有生物学变异进行多变量数据分析。通过气相色谱-飞行时间质谱分析代谢物、二维液相色谱-质谱分析蛋白质以及Northern印迹分析RNA，验证了在单个拟南芥叶片样品中检测到652种代谢物、297种蛋白质和清晰的RNA条带。将不同拟南芥种质重复分析中最丰富的蛋白质和代谢物的一个子集合并，形成一个综合数据集，既可以对不同基因型进行分类，也可以对真实生化网络中蛋白质和代谢物的层次组织进行无偏分析。
