Department of Biochemistry and Food Chemistry, Molecular Plant Biology, University of Turku, Finland.
J Proteome Res. 2010 Nov 5;9(11):5896-912. doi: 10.1021/pr100651w. Epub 2010 Sep 20.
Cyanobacteria developed efficient carbon concentrating mechanisms which significantly improve the photosynthetic performance and survival of cells under limiting CO(2) conditions. Dynamic changes of the Synechocystis proteome to CO(2) limitation were investigated using shotgun LC-MS/MS approach with isobaric tag for relative and absolute quantification (iTRAQ) technique. Synechocystis cells grown at high (3%) CO(2) were shifted to air-level CO(2) followed by protein extraction after 6, 24, and 72 h. About 19% of the cyanobacterial proteome was identified and the expression changes were quantified for 17% of theoretical ORFs. For 76 proteins, up- or down-regulation was found to be significant (more than 1.5 or less than 0.7). Major changes were observed in proteins participating in inorganic carbon uptake, CO(2) fixation, nitrogen transport and assimilation, as well as in the protection of the photosynthetic machinery from excess of light. Further, a number of hypothetical proteins with unknown functions were discovered. In general, the cells appear to acclimate to low CO(2) without a significant stress since the stress-related molecular chaperones were down-regulated and only a minor decline was detected for proteins of phycobilisomes, photosynthetic complexes, and translation machinery. The results of iTRAQ experiment were validated by the Western blot analysis for selected proteins.
蓝藻进化出了高效的碳浓缩机制,这显著提高了细胞在有限 CO₂条件下的光合作用性能和生存能力。本研究采用同位素相对标记与绝对定量技术(iTRAQ)的 shotgun LC-MS/MS 方法,研究了 Synechocystis 蛋白组在 CO₂限制下的动态变化。将在高(3%)CO₂条件下生长的 Synechocystis 细胞转移到空气水平的 CO₂环境中,然后在 6、24 和 72 h 后提取蛋白质。鉴定出约 19%的蓝藻蛋白组,定量了 17%理论 ORFs 的表达变化。对于 76 种蛋白质,发现其上调或下调的幅度都大于 1.5 或小于 0.7。在参与无机碳摄取、CO₂固定、氮运输和同化的蛋白质,以及保护光合作用机器免受过量光的蛋白质中观察到了主要变化。此外,还发现了一些具有未知功能的假设蛋白。总的来说,细胞似乎在没有明显压力的情况下适应了低 CO₂,因为应激相关分子伴侣下调,而藻胆体、光合作用复合物和翻译机制的蛋白质仅略有下降。通过对选定蛋白质的 Western blot 分析验证了 iTRAQ 实验的结果。