Department of Molecular Structural Biology, Max Planck Institute of Biochemistry, Am Klopferspitz 18, D-82152 Martinsried, Germany.
J Proteome Res. 2010 Sep 3;9(9):4839-50. doi: 10.1021/pr100567u.
A comparative proteome and transcriptome analysis of Thermoplasma acidophilum cultured under aerobic and anaerobic conditions has been performed. One-thousand twenty-five proteins were identified covering 88% of the cytosolic proteome. Using a label-free quantitation method, we found that approximately one-quarter of the identified proteome (263 proteins) were significantly induced (>2 fold) under anaerobic conditions. Thirty-nine macromolecular complexes were identified, of which 28 were quantified and 15 were regulated under anaerobiosis. In parallel, a whole genome cDNA microarray analysis was performed showing that the expression levels of 445 genes were influenced by the absence of oxygen. Interestingly, more than 40% of the membrane protein-encoding genes (145 out of 335 ORFs) were up- or down-regulated at the mRNA level. Many of these proteins are functionally associated with extracellular protein or peptide degradation or ion and amino acid transport. Comparison of the transcriptome and proteome showed only a weak positive correlation between mRNA and protein expression changes, which is indicative of extensive post-transcriptional regulatory mechanisms in T. acidophilum. Integration of transcriptomics and proteomics data generated hypotheses for physiological adaptations of the cells to anaerobiosis, and the quantitative proteomics data together with quantitative analysis of protein complexes provide a platform for correlation of MS-based proteomics studies with cryo-electron tomography-based visual proteomics approaches.
已对嗜酸性热原体在有氧和无氧条件下培养的比较蛋白质组和转录组进行了分析。鉴定了 1025 种蛋白质,涵盖了细胞质蛋白质组的 88%。使用无标记定量方法,我们发现约四分之一的鉴定蛋白质组(263 种蛋白质)在厌氧条件下显著诱导(>2 倍)。鉴定了 39 个大分子复合物,其中 28 个进行了定量分析,15 个在厌氧条件下受到调节。同时,进行了全基因组 cDNA 微阵列分析,结果显示 445 个基因的表达水平受到缺氧的影响。有趣的是,超过 40%的膜蛋白编码基因(335 个 ORF 中的 145 个)在 mRNA 水平上上调或下调。这些蛋白质中的许多与细胞外蛋白质或肽降解或离子和氨基酸转运功能相关。转录组和蛋白质组的比较仅显示 mRNA 和蛋白质表达变化之间的弱正相关性,这表明嗜酸性热原体中存在广泛的转录后调控机制。转录组学和蛋白质组学数据的整合为细胞对厌氧条件的生理适应提供了假设,并且定量蛋白质组学数据以及蛋白质复合物的定量分析为基于 MS 的蛋白质组学研究与基于冷冻电子断层扫描的可视化蛋白质组学方法的相关性提供了平台。