Protein transport across and into cell membranes in bacteria and archaea

Yuan, Jijun; Zweers, Jessica C.; Dijl, Jan Maarten van; Dalbey, Ross E.

doi:10.1007/s00018-009-0160-x

Cited by 84 publications

(76 citation statements)

References 226 publications

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“…This protein export system recognizes substrates bearing cleavable N-terminal signal peptides, and transports them through the Abbreviations DCW, dry cell weight; EM, electron microscopy; GFP, green fluorescent protein; IPTG, isopropyl thio-b-D-galactoside; Sec, general secretory; Tat, twin-arginine translocation; WT, wild-type. membrane-bound translocase in an unfolded form (for a review, see [3]). …”

Section: Introductionmentioning

confidence: 99%

High‐level secretion of a recombinant protein to the culture medium with a Bacillus subtilis twin‐arginine translocation system in Escherichia coli

et al. 2013

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The twin‐arginine translocation (Tat) system transports folded proteins across the plasma membrane in bacteria, and heterologous proteins can be exported by this pathway if a Tat‐type signal peptide is present at the N‐terminus. The system thus has potential for biopharmaceutical production in Escherichia coli, where export to the periplasm is often a favoured approach. Previous studies have shown that E. coli cells can export high levels of protein by the Tat pathway, and the protein product accummulates almost exclusively in the periplasm. In this study, we analysed E. coli cells that express the Bacillus subtilis TatAdCd system in place of the native TatABC system. We show that a heterologous model protein, comprising the TorA signal peptide linked to green fluorescent protein (TorA–GFP), is efficiently exported by the TatAdCd system. However, whereas the GFP is exported initially to the periplasm during batch fermentation, the mature protein is increasingly found in the extracellular culture medium. By the end of a 16‐h fermentation, ~ 90% of exported GFP is present in the medium as active mature protein. The total protein profiles of the medium and periplasm are essentially identical, confirming that the outer membrane becomes leaky during the fermentation process. The cells are otherwise intact, and there is no large‐scale release of cytoplasmic contents. Export levels are relatively high, with ~ 0.35 g GFP·L−1 culture present in the medium. This system thus offers a means of producing recombinant protein in E. coli and harvesting directly from the medium, with potential advantages in terms of ease of purification and downstream processing.

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Section: Introductionmentioning

confidence: 99%

High‐level secretion of a recombinant protein to the culture medium with a Bacillus subtilis twin‐arginine translocation system in Escherichia coli

et al. 2013

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“…Targeting of these proteins in eubacteria and archaea occurs through the wellcharacterized Sec and Tat translocon pathways (10). Protein substrates for the Sec and Tat systems were shown to be distinguished by specific N-terminal signal sequences (11).…”

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confidence: 99%

Deletion of Synechocystis sp. PCC 6803 Leader Peptidase LepB1 Affects Photosynthetic Complexes and Respiration

Zhang

Selão

Pisareva

et al. 2013

Molecular & Cellular Proteomics

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The cyanobacterium Synechocystis sp. PCC 6803 possesses two leader peptidases, LepB1 (Sll0716) and LepB2 (Slr1377), responsible for the processing of signal peptide-containing proteins. Deletion of the gene for LepB1 results in an inability to grow photoautotrophically and an extreme light sensitivity. Here we show, using a combination of Blue Native/SDS-PAGE, Western blotting and iTRAQ analysis, that lack of LepB1 strongly affects the cell's ability to accumulate wild-type levels of both photosystem I (PSI) and cytochrome (Cyt) b 6 f complexes. The impaired assembly of PSI and Cyt b 6 f is considered to be caused by the no or slow processing of the integral subunits PsaF and Cyt f respectively. In particular, PsaF, one of the PSI subunits, was found incorporated into PSI in its unprocessed form, which could influence the assembly and/or stability of PSI. In contrast to these results, we found the amount of assembled photosystem II (PSII) unchanged, despite a slower processing of PsbO. Thus, imbalance in the ratios of PSI and Cyt b 6 f to photosystem II leads to an imbalanced photosynthetic electron flow upand down-stream of the plastoquinone pool, resulting in the observed light sensitivity of the mutant. We conclude that LepB1 is the natural leader peptidase for PsaF, PsbO, and Cyt f. The maturation of PsbO and Cyt f can be partially performed by LepB2, whereas PsaF processing is completely dependent on LepB1. iTRAQ analysis also revealed a number of indirect effects accompanying the mutation, primarily a strong induction of the CydAB oxidase as well as a significant decrease in phycobiliproteins and chlorophyll/heme biosynthesis enzymes. Molecular

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“…YidC is an essential E. coli IMP that plays an important but poorly defined role in the biogenesis of IMPs (18,52). So far, targeted approaches have revealed a plethora of YidC functions that seem almost too varied to be combined in one protein.…”

Section: Discussionmentioning

confidence: 99%

“…These inner membrane proteins (IMPs) first are targeted to and then inserted into the IM, after which they fold and, if required, assemble into oligomeric complexes (51). The Sec translocon is the main protein-conducting channel used to insert IMPs (18,52). The translocon consists of the heterotrimeric channel complex SecYEG and the accessory components YidC and SecDF YajC.…”

mentioning

confidence: 99%

Activators of the Glutamate-Dependent Acid Resistance System Alleviate Deleterious Effects of YidC Depletion inEscherichia coli

Yu¹,

Bekker

Tramonti

et al. 2011

J Bacteriol

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The function of the essential inner membrane protein (IMP) YidC in Escherichia coli has been studied for a limited number of model IMPs and primarily using targeted approaches. These studies suggested that YidC acts at the level of insertion, folding, and quality control of IMPs, both in the context of the Sec translocon and as a separate entity. To further our understanding of YidC's role in IMP biogenesis, we screened a random overexpression library for factors that rescued the growth of cells upon YidC depletion. We found that the overexpression of the GadX and GadY regulators of the glutamate-dependent acid resistance system complemented the growth defect of YidC-depleted cells. Evidence is presented that GadXY overexpression counteracts the deleterious effects of YidC depletion on at least two fronts. First, GadXY prepares the cells for the decrease in respiratory capacity upon the depletion of YidC. Most likely, GadXY-regulated processes reduce the drop in proton-motive force that impairs the fitness of YidC-depleted cells. Second, in GadXY-overproducing cells increased levels of the general chaperone GroEL cofractionate with the inner membranes, which may help to keep newly synthesized inner membrane proteins in an insertion-competent state when YidC levels are limiting.

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Protein transport across and into cell membranes in bacteria and archaea

Cited by 84 publications

References 226 publications

High‐level secretion of a recombinant protein to the culture medium with a Bacillus subtilis twin‐arginine translocation system in Escherichia coli

High‐level secretion of a recombinant protein to the culture medium with a Bacillus subtilis twin‐arginine translocation system in Escherichia coli

Deletion of Synechocystis sp. PCC 6803 Leader Peptidase LepB1 Affects Photosynthetic Complexes and Respiration

Activators of the Glutamate-Dependent Acid Resistance System Alleviate Deleterious Effects of YidC Depletion inEscherichia coli

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