Fundamental & Computational Sciences Directorate, Pacific Northwest National Laboratory, Richland, WA, 99352, USA.
Environ Microbiol Rep. 2014 Dec;6(6):776-85. doi: 10.1111/1758-2229.12204. Epub 2014 Sep 24.
The multi-heme, outer membrane c-type cytochrome (c-Cyt) OmcB of Geobacter sulfurreducens was previously proposed to mediate electron transfer across the outer membrane. However, the underlying mechanism has remained uncharacterized. In G. sulfurreducens, the omcB gene is part of two tandem four-gene clusters, each is predicted to encode a transcriptional factor (OrfR/OrfS), a porin-like outer membrane protein (OmbB/OmbC), a periplasmic c-type cytochrome (OmaB/OmaC) and an outer membrane c-Cyt (OmcB/OmcC) respectively. Here, we showed that OmbB/OmbC, OmaB/OmaC and OmcB/OmcC of G. sulfurreducens PCA formed the porin-cytochrome (Pcc) protein complexes, which were involved in transferring electrons across the outer membrane. The isolated Pcc protein complexes reconstituted in proteoliposomes transferred electrons from reduced methyl viologen across the lipid bilayer of liposomes to Fe(III)-citrate and ferrihydrite. The pcc clusters were found in all eight sequenced Geobacter and 11 other bacterial genomes from six different phyla, demonstrating a widespread distribution of Pcc protein complexes in phylogenetically diverse bacteria. Deletion of ombB-omaB-omcB-orfS-ombC-omaC-omcC gene clusters had no impact on the growth of G. sulfurreducens PCA with fumarate but diminished the ability of G. sulfurreducens PCA to reduce Fe(III)-citrate and ferrihydrite. Complementation with the ombB-omaB-omcB gene cluster restored the ability of G. sulfurreducens PCA to reduce Fe(III)-citrate and ferrihydrite.
先前有研究提出,脱硫杆菌的多血红素外膜 c 型细胞色素(c-Cyt)OmcB 介导了电子在外膜间的传递。然而,其潜在的机制仍未被阐明。在脱硫杆菌中,omcB 基因是两个串联的四基因簇的一部分,每个基因簇都预测编码一个转录因子(OrfR/OrfS)、一个孔蛋白样外膜蛋白(OmbB/OmbC)、一个周质 c 型细胞色素(OmaB/OmaC)和一个外膜 c-Cyt(OmcB/OmcC)。在这里,我们发现脱硫杆菌 PCA 的 OmbB/OmbC、OmaB/OmaC 和 OmcB/OmcC 形成了孔-细胞色素(Pcc)蛋白复合物,该复合物参与了电子在外膜间的传递。在蛋白脂质体中分离出的 Pcc 蛋白复合物将还原的甲紫精中的电子从脂质体的脂质双层中转移到 Fe(III)-柠檬酸盐和水铁矿。pcc 簇在所有八个测序的 Geobacter 和来自六个不同门的 11 个其他细菌基因组中都有发现,这表明 Pcc 蛋白复合物在系统发育上多样化的细菌中广泛分布。缺失 ombB-omaB-omcB-orfS-ombC-omaC-omcC 基因簇对脱硫杆菌 PCA 利用富马酸盐的生长没有影响,但降低了脱硫杆菌 PCA 还原 Fe(III)-柠檬酸盐和水铁矿的能力。ompB-omaB-omcB 基因簇的互补恢复了脱硫杆菌 PCA 还原 Fe(III)-柠檬酸盐和水铁矿的能力。
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