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通过瑞士卡达格诺湖水柱对一种绿色硫细菌的比较蛋白质组学和活性研究。

Comparative proteomics and activity of a green sulfur bacterium through the water column of Lake Cadagno, Switzerland.

机构信息

Department of Biology and Nordic Center for Earth EvolutionDepartment of Biochemistry and Molecular Biology, University of Southern Denmark, DK-5230 Odense M, Denmark.Department of Biology, University of Copenhagen, DK-2200 Copenhagen N, Denmark.Cantonal Institute of Microbiology, CH-6500 Bellinzona and Microbial Ecology and Microbiology Unit, Plant Biology Department, University of Geneva, CH-1211 Geneva, Switzerland.Alpine Biology Center Foundation Piora, Quinto, Switzerland.

出版信息

Environ Microbiol. 2011 Jan;13(1):203-215. doi: 10.1111/j.1462-2920.2010.02321.x. Epub 2010 Aug 20.

Abstract

Primary production in the meromictic Lake Cadagno, Switzerland, is dominated by anoxygenic photosynthesis. The green sulfur bacterium Chlorobium clathratiforme is the dominant phototrophic organism in the lake, comprising more than half of the bacterial population, and its biomass increases 3.8-fold over the summer. Cells from four positions in the water column were used for comparative analysis of the Chl. clathratiforme proteome in order to investigate changes in protein composition in response to the chemical and physical gradient in their environment, with special focus on how the bacteria survive in the dark. Although metagenomic data are not available for Lake Cadagno, proteome analysis was possible based on the completely sequenced genome of an isolated strain of Chl. clathratiforme. Using LC-MS/MS we identified 1321 Chl. clathratiforme proteins in Lake Cadagno and quantitatively compared 621 of these in the four samples. Our results showed that compared with cells obtained from the photic zone, cells collected from the dark part of the water column had the same expression level of key enzymes involved in carbon metabolism and photosynthetic light harvesting. However, most proteins participating in nitrogen and sulfur metabolism were twofold less abundant in the dark. From the proteome analysis we were able to show that Chl. clathratiforme in the photic zone contains enzymes for fixation of N(2) and the complete oxidation of sulfide to sulfate while these processes are probably not active in the dark. Instead we propose that Chl. clathratiforme cells in the dark part of the water column obtain energy for maintenance from the fermentation of polyglucose. Based on the observed protein compositions we have constructed possible pathways for C, N and S metabolism in Chl. clathratiforme.

摘要

瑞士中营养分层湖卡达戈的初级生产力主要由厌氧光合作用主导。绿硫细菌绿菌属(Chlorobium clathratiforme)是湖中占优势的光合生物,占细菌总数的一半以上,其生物量在夏季增加了 3.8 倍。从水柱的四个位置采集细胞,用于比较分析 Chl. clathratiforme 的蛋白质组,以研究蛋白质组成的变化以响应其环境中的化学和物理梯度,特别关注细菌如何在黑暗中生存。尽管卡达戈湖没有宏基因组数据,但基于分离的绿菌属(Chlorobium clathratiforme)菌株的完整测序基因组,可以进行蛋白质组分析。使用 LC-MS/MS,我们在卡达戈湖中鉴定出 1321 种绿菌属(Chlorobium clathratiforme)蛋白,并对这四种样本中的 621 种蛋白进行了定量比较。我们的结果表明,与从光区获得的细胞相比,从水柱的黑暗部分采集的细胞具有相同水平的参与碳代谢和光合作用光捕获的关键酶。然而,在黑暗中,参与氮和硫代谢的大多数蛋白质的丰度减少了两倍。从蛋白质组分析中,我们能够表明在光区的绿菌属(Chlorobium clathratiforme)含有固定 N(2)和将硫化物完全氧化为硫酸盐的酶,而这些过程在黑暗中可能不活跃。相反,我们提出在水柱的黑暗部分的绿菌属(Chlorobium clathratiforme)细胞可能从多糖发酵中获取维持所需的能量。根据观察到的蛋白质组成,我们构建了绿菌属(Chlorobium clathratiforme)中 C、N 和 S 代谢的可能途径。

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