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在以富马酸盐或柠檬酸铁为碳源生长的金属还原菌嗜硫产甲烷杆菌PCA中的差异蛋白质表达

Differential protein expression in the metal-reducing bacterium Geobacter sulfurreducens strain PCA grown with fumarate or ferric citrate.

作者信息

Khare Tripti, Esteve-Núñez Abraham, Nevin Kelly P, Zhu Wenhong, Yates John R, Lovley Derek, Giometti Carol S

机构信息

Argonne National Laboratory, Argonne, Illinois 60439, USA.

出版信息

Proteomics. 2006 Jan;6(2):632-40. doi: 10.1002/pmic.200500137.

Abstract

Geobacter sulfurreducens, generally considered to be a strict anaerobe, is a predominant microbe in subsurface environments, where it utilizes available metals as electron acceptors. To better understand the metabolic processes involved in the metal-reduction capability of this microbe, the proteins expressed by cells grown anaerobically with either fumarate or ferric citrate as electron acceptor were compared. Proteins were separated by 2-DE under denaturing or nondenaturing conditions, and proteins varying in abundance with a high level of statistical significance (p<0.0001) were identified by peptide mass analysis. Denaturing 2-DE revealed significant differences in the relative abundance of the membrane proteins OmpA and peptidoglycan-associated lipoprotein, several metabolic enzymes, and, in addition, superoxide dismutase and rubredoxin oxidoreductase. Nondenaturing 2-DE revealed elevated catalase in cells grown with ferric citrate. These results suggest that, in addition to adjustments in membrane transport and specific metabolic pathways in response to these two different electron acceptors, distinct differences exist in the oxidative environment within the cell when fumarate or soluble ferric citrate is provided as electron acceptor. Although an anaerobe, G. sulfurreducens appears to have alternate mechanisms for dealing with reactive oxygen species in response to increased intracellular soluble iron.

摘要

硫还原地杆菌通常被认为是一种严格厌氧菌,是地下环境中的优势微生物,在该环境中它利用可利用的金属作为电子受体。为了更好地理解这种微生物的金属还原能力所涉及的代谢过程,对以富马酸或柠檬酸铁作为电子受体进行厌氧培养的细胞所表达的蛋白质进行了比较。蛋白质在变性或非变性条件下通过双向电泳进行分离,并通过肽质量分析鉴定出丰度有显著差异且具有高度统计学意义(p<0.0001)的蛋白质。变性双向电泳显示,膜蛋白OmpA和肽聚糖相关脂蛋白、几种代谢酶以及超氧化物歧化酶和红素氧化还原酶的相对丰度存在显著差异。非变性双向电泳显示,以柠檬酸铁培养的细胞中过氧化氢酶水平升高。这些结果表明,除了针对这两种不同电子受体对膜转运和特定代谢途径进行调整外,当以富马酸或可溶性柠檬酸铁作为电子受体时,细胞内的氧化环境存在明显差异。尽管硫还原地杆菌是厌氧菌,但它似乎具有应对细胞内可溶性铁增加时产生的活性氧的替代机制。

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