Schmidt Michael W, Houseman Andres, Ivanov Alexander R, Wolf Dieter A
NIEHS Center for Environmental Health Proteomics Facility, Harvard School of Public Health, Boston, MA 02115, USA.
Mol Syst Biol. 2007;3:79. doi: 10.1038/msb4100117. Epub 2007 Feb 13.
The fission yeast Schizosaccharomyces pombe is a widely used model organism to study basic mechanisms of eukaryotic biology, but unlike other model organisms, its proteome remains largely uncharacterized. Using a shotgun proteomics approach based on multidimensional prefractionation and tandem mass spectrometry, we have detected approximately 30% of the theoretical fission yeast proteome. Applying statistical modelling to normalize spectral counts to the number of predicted tryptic peptides, we have performed label-free quantification of 1465 proteins. The fission yeast protein data showed considerable correlations with mRNA levels and with the abundance of orthologous proteins in budding yeast. Functional pathway analysis indicated that the mRNA-protein correlation is strong for proteins involved in signalling and metabolic processes, but increasingly discordant for components of protein complexes, which clustered in groups with similar mRNA-protein ratios. Self-organizing map clustering of large-scale protein and mRNA data from fission and budding yeast revealed coordinate but not always concordant expression of components of functional pathways and protein complexes. This finding reaffirms at the protein level the considerable divergence in gene expression patterns of the two model organisms that was noticed in previous transcriptomic studies.
裂殖酵母粟酒裂殖酵母是一种广泛用于研究真核生物生物学基本机制的模式生物,但与其他模式生物不同的是,其蛋白质组在很大程度上仍未得到表征。使用基于多维预分级和串联质谱的鸟枪法蛋白质组学方法,我们检测到了约30%的理论裂殖酵母蛋白质组。应用统计模型将光谱计数归一化为预测的胰蛋白酶肽数量,我们对1465种蛋白质进行了无标记定量。裂殖酵母蛋白质数据与mRNA水平以及与芽殖酵母中直系同源蛋白质的丰度显示出相当的相关性。功能途径分析表明,对于参与信号传导和代谢过程的蛋白质,mRNA与蛋白质的相关性很强,但对于蛋白质复合物的成分,这种相关性越来越不一致,这些成分按具有相似mRNA与蛋白质比率的组聚集。来自裂殖酵母和芽殖酵母的大规模蛋白质和mRNA数据的自组织映射聚类揭示了功能途径和蛋白质复合物成分的协同但并非总是一致的表达。这一发现再次在蛋白质水平上证实了在先前的转录组学研究中注意到的两种模式生物在基因表达模式上的显著差异。