Kurian Dominic, Jansèn Tove, Mäenpää Pirkko
Laboratory of Plant Physiology and Molecular Biology, Department of Biology, University of Turku, Turku, Finland.
Proteomics. 2006 Mar;6(5):1483-94. doi: 10.1002/pmic.200500413.
To provide an insight into the heterotrophic metabolism of cyanobacteria, a proteomic approach has been employed with the model organism Synechocystis sp. PCC 6803. The soluble proteins from Synechocystis grown under photoautotrophic and light-activated heterotrophic conditions were separated by 2-DE and identified by MALDI-MS or LC-MS/MS analysis. 2-DE gels made using narrow- and micro-range IPG strips allowed quantitative comparison of more than 900 spots. Out of 67 abundant protein spots identified, 13 spots were increased and 9 decreased under heterotrophy, representing all the major fold changes. Proteomic alterations and activity levels of selected enzymes indicate a shift in the central carbon metabolism in response to trophic change. The significant reduction in light-saturated rate of photosynthesis as well as in the expression levels of rubisco and CO(2)-concentrating mechanism proteins under heterotrophy indicates the down-regulation of the photosynthetic machinery. Alterations in the expression level of proteins involved in carbon utilization pathways refer to enhanced glycolysis, oxidative pentose phosphate pathway as well as tricarboxylic acid cycle under heterotrophy. Proteomic evidences also suggest an enhanced biosynthesis of amino acids such as histidine and serine during heterotrophic growth.
为深入了解蓝藻的异养代谢,采用蛋白质组学方法对模式生物集胞藻PCC 6803进行研究。将集胞藻在光合自养和光激活异养条件下生长产生的可溶性蛋白质通过二维电泳(2-DE)分离,并通过基质辅助激光解吸电离质谱(MALDI-MS)或液相色谱-串联质谱(LC-MS/MS)分析进行鉴定。使用窄范围和微范围的固相pH梯度(IPG)条制作的二维电泳凝胶能够对900多个斑点进行定量比较。在鉴定出的67个丰富蛋白质斑点中,有13个斑点在异养条件下增加,9个斑点减少,代表了所有主要的倍数变化。蛋白质组学变化和所选酶的活性水平表明,中央碳代谢响应营养变化发生了转变。异养条件下光合光饱和速率以及核酮糖-1,5-二磷酸羧化酶/加氧酶(rubisco)和二氧化碳浓缩机制蛋白表达水平的显著降低表明光合机构受到下调。参与碳利用途径的蛋白质表达水平的变化表明异养条件下糖酵解、氧化戊糖磷酸途径以及三羧酸循环增强。蛋白质组学证据还表明,在异养生长期间,组氨酸和丝氨酸等氨基酸的生物合成增强。