Bosch Gundula, Wang Tiansong, Latypova Ekaterina, Kalyuzhnaya Marina G, Hackett Murray, Chistoserdova Ludmila
Department of Chemical Engineering, University of Washington, Seattle, WA 98195, USA.
Department of Microbiology, University of Washington, Seattle, WA 98195, USA.
Microbiology (Reading). 2009 Apr;155(Pt 4):1103-1110. doi: 10.1099/mic.0.024968-0.
While the shotgun proteomics approach is gaining momentum in understanding microbial physiology, it remains limited by the paucity of high-quality genomic data, especially when it comes to poorly characterized newly identified phyla. At the same time, large-scale metagenomic sequencing projects produce datasets representing genomes of a variety of environmental microbes, although with lower sequence coverage and sequence quality. In this work we tested the utility of a metagenomic dataset enriched in sequences of environmental strains of Methylotenera mobilis, to assess the protein profile of a laboratory-cultivated strain, M. mobilis JLW8, as a proof of principle. We demonstrate that a large portion of the proteome predicted from the metagenomic sequence (approx. 20 %) could be identified with high confidence (three or more peptide sequences), thus gaining insights into the physiology of this bacterium, which represents a new genus within the family Methylophilaceae.
虽然鸟枪法蛋白质组学方法在理解微生物生理学方面正获得越来越多的关注,但它仍然受到高质量基因组数据匮乏的限制,特别是在涉及特征描述不足的新鉴定门类时。与此同时,大规模宏基因组测序项目产生了代表各种环境微生物基因组的数据集,尽管其序列覆盖度和序列质量较低。在这项工作中,我们测试了一个富含活动甲基弯曲菌环境菌株序列的宏基因组数据集的效用,以评估实验室培养菌株活动甲基弯曲菌JLW8的蛋白质谱,作为原理验证。我们证明,从宏基因组序列预测的很大一部分蛋白质组(约20%)可以被高置信度地鉴定(三个或更多肽序列),从而深入了解这种细菌的生理学,它代表嗜甲基菌科中的一个新属。