Proteomics with a pinch of salt: a cyanobacterial perspective.
Proteomics with a pinch of salt: a cyanobacterial perspective.
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DOI：
10.1186/1746-1448-4-1
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发表时间：
2008-04-15
期刊：
影响因子：
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通讯作者：
Biggs CA
中科院分区：
文献类型：
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作者：
Pandhal J;Wright PC;Biggs CA
蓝藻是古老的生命形式，已经适应了各种极端环境，包括高盐度。生物化学、生理学和遗传学研究有助于揭示它们潜在的生存机制，最近的研究表明，蛋白质组学有可能进一步加深我们对它们的整体理解。到目前为止，大多数与盐相关的蓝藻蛋白质组研究都使用了模式生物聚球藻的凝胶电泳法。PCC6803。此外，关注的焦点是不同细胞隔间内2-4%w/v的氯化钠浓度。在此条件下，聚球藻Sp.PCC6803通过合成一般和特定的胁迫蛋白，改变胞外层的蛋白质组成，改变复杂的中央中介通路的调控，来响应和适应盐胁迫。通过不相关的转录水平数据和蛋白质异构体的鉴定，也可以预测转录后控制。在本文中，我们还回顾了蛋白质组数据的技术发展，重点是提高蛋白质组数据的质量和数量，克服盐对样品制备和分析的不利影响。无凝胶方法的发展包括蛋白质和多肽的分级工作流程，这可以增加蛋白质组的覆盖率(聚球藻中的20%。PCC6803)。定量技术在准确性方面也有所提高，使定量的可信度接近甚至超过转录组技术(优于差异表达1.5倍)。此外，体内代谢标记和从头蛋白质测序软件提高了将蛋白质组学应用于未测序的环境分离株的能力。这篇综述中使用的例子是从撒哈拉盐湖中分离出来的蓝藻。
Cyanobacteria are ancient life forms and have adapted to a variety of extreme environments, including high salinity. Biochemical, physiological and genetic studies have contributed to uncovering their underlying survival mechanisms, and as recent studies demonstrate, proteomics has the potential to increase our overall understanding further. To date, most salt-related cyanobacterial proteomic studies have utilised gel electrophoresis with the model organism Synechocystis sp. PCC6803. Moreover, focus has been on 2–4% w/v NaCl concentrations within different cellular compartments. Under these conditions, Synechocystis sp. PCC6803 was found to respond and adapt to salt stress through synthesis of general and specific stress proteins, altering the protein composition of extracellular layers, and re-directing control of complex central intermediary pathways. Post-transcriptional control was also predicted through non-correlating transcript level data and identification of protein isoforms. In this paper, we also review technical developments with emphasis on improving the quality and quantity of proteomic data and overcoming the detrimental effects of salt on sample preparation and analysis. Developments in gel-free methods include protein and peptide fractionation workflows, which can increase coverage of the proteome (20% in Synechocystis sp. PCC6803). Quantitative techniques have also improved in accuracy, resulting in confidence in quantitation approaching or even surpassing that seen in transcriptomic techniques (better than 1.5-fold in differential expression). Furthermore, in vivo metabolic labelling and de novo protein sequencing software have improved the ability to apply proteomics to unsequenced environmental isolates. The example used in this review is a cyanobacterium isolated from a Saharan salt lake.