两种金属还原剂的故事:硫还原地杆菌PCA和希瓦氏菌mr1的比较蛋白质组分析。

Methods of biochemical analysis Pub Date : 2006-01-01
Carol S Giometti
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引用次数: 0

摘要

硫还原地杆菌(Geobacter sulphur reducens)和臭杆菌(s.o.o.idensis)因其在生物修复方面的潜在应用而成为人们研究的热点。它们的蛋白质组的特征,与它们的基因组序列、转录组和代谢组的分析同时进行,为它们的相似性和差异性提供了有价值的见解。这两种金属还原微生物的蛋白质组学的主要目标是表征它们的c型细胞色素。发现它们的c型细胞色素的完整补充和描述什么生长条件触发它们的表达是利用它们的生物修复潜力的核心。迄今为止的蛋白质组学分析表明,G. sulphreducens和S. oneidensis的大多数c型细胞色素在它们的外膜上都有共同的位置。然而,G.硫还原菌的c型细胞色素可溶性较低,因此比S. oneidensis表达的c型细胞色素更难从膜中分离出来。大多数硫还原g的c型细胞色素也不同于草的c型细胞色素,它们具有更高的等电点,大多数高于pH 8.0。这些溶解度和等电点的特征可能是相关的,并且可能表明这两种微生物在金属还原策略上的潜在功能差异。在撰写本文时,可获得的G.硫还原菌和S. oneidensis的全球蛋白质组学结果主要是2DE分析结合LC-MS/MS对凝胶内消化的色氨酸肽进行蛋白质鉴定的结果,代表了考马斯蓝或硝酸银染色检测到的最丰富的蛋白质。目前,一些利用2D-LC-MS/MS方法的互补工作正在进行中,有望在不久的将来为这些微生物提供更完整的蛋白质清单。随着这些数据的补充,代谢过程的复杂网络、蛋白质合成和蛋白质功能的调节、营养物质的运输和信号转导将得到阐明。现有的蛋白质组学工具将与蛋白质芯片和噬菌体展示等新方法相辅相成,以进一步表征这些微生物系统。在不太遥远的将来,最终结果将是在各种环境条件下,在其自然栖息地中硫还原G.和S. oneidensis行为的预测模型。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Tale of two metal reducers: comparative proteome analysis of Geobacter sulferreducens PCA and Shewanella oneidensis MR-1.

Geobacter sulfurreducens and S. oneidensis are the subjects of intense research efforts due to their potential applications to bioremediation. The characterization of their proteomes, being done in parallel with the analysis of their genome sequences, transcriptomes, and metabolomes, is providing valuable insights to both their similarities and their differences. A primary target of interest in the proteomes of both of these metal-reducing microbes is the characterization of their c-type cytochromes. The discovery of their full compliment of c-type cytochromes and the description of what growth conditions trigger their expression is central to harnessing their bioremediation potential. Proteome analyses thus far show that both G. sulfurreducens and S. oneidensis share the common location of a majority of their c-type cytochromes in their outer membranes. The c-type cytochromes of G. sulfurreducens, however, appear to be less soluble and therefore more difficult to isolate from the membranes than those expressed by S. oneidensis. The majority of the G. sulfurreducens c-type cytochromes also differ from those of S. oneidensis in that they have higher isoelectric points, most higher than pH 8.0. These characteristics of solubility and isoelectric point could be related and could indicate an underlying functional difference in the strategy for metal reduction between these two microbes. The global proteome results available for G. sulfurreducens and S. oneidensis at the time of this writing are primarily the result of 2DE analysis coupled to protein identification by LC-MS/MS of tryptic peptides from in-gel digests and represent the most abundant proteins detected by Coomassie blue or silver nitrate staining. Currently, several complimentary efforts utilising the 2D-LC-MS/MS approaches are in progress, promising a more complete protein inventory for these microbes in the near future. As these data are added to those already available, the intricate network of metabolic processes, regulation of protein synthesis and protein function, transport of nutrients, and signal transduction will be elucidated. The existing tools of proteomics will be complimented with newer methods such as protein chips and phage display to further characterize these microbial systems. The end result, in the not too distant future, will be predictive models of G. sulfurreducens and S. oneidensis behavior in their natural habitats under a variety of environmental conditions.

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