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略带保留的蛋白质组学:从蓝细菌角度看

Proteomics with a pinch of salt: a cyanobacterial perspective.

作者信息

Pandhal Jagroop, Wright Phillip C, Biggs Catherine A

机构信息

Department of Chemical and Process Engineering, The University of Sheffield, Sheffield, UK.

出版信息

Saline Syst. 2008 Apr 15;4:1. doi: 10.1186/1746-1448-4-1.

Abstract

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.

摘要

蓝藻是古老的生命形式,已经适应了包括高盐度在内的各种极端环境。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,而且正如最近的研究所表明的,蛋白质组学有潜力进一步增进我们的整体理解。迄今为止,大多数与盐相关的蓝藻蛋白质组学研究都利用模式生物聚球藻属PCC6803进行凝胶电泳分析。此外,研究重点一直是不同细胞区室内2-4% w/v的氯化钠浓度。在这些条件下,发现聚球藻属PCC6803通过合成一般和特定的应激蛋白、改变细胞外层的蛋白质组成以及重新引导复杂的中央中间代谢途径的控制来响应和适应盐胁迫。转录后调控也通过不相关的转录水平数据和蛋白质异构体的鉴定而得到预测。在本文中,我们还综述了技术发展,重点是提高蛋白质组学数据的质量和数量,以及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肽段分级分离工作流程,这可以增加蛋白质组的覆盖率(在聚球藻属PCC6803中提高20%)。定量技术的准确性也有所提高,使得定量的可信度接近甚至超过转录组学技术(差异表达时优于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环境分离株的能力。本综述中使用的例子是一种从撒哈拉盐湖分离出的蓝藻。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/681d/2386806/a07ff6b4d582/1746-1448-4-1-1.jpg

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