Structure analysis of the protein translocating channel TatA in membranes using a multi-construct approach

Lange, Christian; Müller, Sebastian; Walther, Torsten H.; Bürck, Jochen; Ulrich, Anne S.

doi:10.1016/j.bbamem.2007.06.021

Cited by 42 publications

(47 citation statements)

References 28 publications

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“…The low transport activity of our TatA-YFP fusion is consistent with other studies that used fusions to the C terminus of TatA (refs 38 and 39 and T.P., unpublished observations) and occurs even though the last Ϸ50 aa of TatA have a random coil structure (12,40). Steric hindrance to the movement of the substrate is a likely explanation for this phenomenon.…”

Section: Discussionsupporting

confidence: 90%

Variable stoichiometry of the TatA component of the twin-arginine protein transport system observed by in vivo single-molecule imaging

Leake

Greene

Godun

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

175

243

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The twin-arginine translocation (Tat) system transports folded proteins across the bacterial cytoplasmic membrane and the thylakoid membrane of plant chloroplasts. The essential components of the Tat pathway are the membrane proteins TatA, TatB, and TatC. TatA is thought to form the protein translocating element of the Tat system. Current models for Tat transport make predictions about the oligomeric state of TatA and whether, and how, this state changes during the transport cycle. We determined the oligomeric state of TatA

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Section: Discussionsupporting

confidence: 90%

Variable stoichiometry of the TatA component of the twin-arginine protein transport system observed by in vivo single-molecule imaging

Leake

Greene

Godun

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

175

243

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“…1C and Refs. [55][56][57][58]. Furthermore, functional analyses suggested that the conserved hinge region connecting both helices might be of structural importance for TatAd (50).…”

Section: Discussionmentioning

confidence: 99%

Salt Sensitivity of Minimal Twin Arginine Translocases

Ploeg

Barnett

Vasisht

et al. 2011

Journal of Biological Chemistry

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Background: Minimal twin arginine preprotein translocases consist of TatA and TatC subunits. Results: Here we show that various mutated and wild-type minimal Tat translocases are salt-sensitive. Conclusion: Salt-sensitive electrostatic interactions seem to have critical roles in the preprotein translocation activity of minimal Tat translocases. Significance: Our results represent first experimental evidence for an important role of salt-sensitive electrostatic interactions in Tat translocation.

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“…However, currently no detailed structures of the TAT apparatus or of any of its individual proteins (i.e., TatA, -B, or -C) currently exist for molecular modeling with promising lead compounds (32,43,67). Additionally, although an in vitro based assay of TAT function has been described for E. coli TAT, it has not yet been used for evaluating small-molecule TAT inhibitors, and it has not yet been developed for the P. aeruginosa TAT system (70).…”

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confidence: 99%

Identification and Evaluation of Twin-Arginine Translocase Inhibitors

Vasil

Tomaras

Pritchard

2012

Antimicrob Agents Chemother

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The twin-arginine translocase (TAT) in some bacterial pathogens, including Pseudomonas aeruginosa, Burkholderia pseudomallei, and Mycobacterium tuberculosis, contributes to pathogenesis by translocating extracellular virulence determinants across the inner membrane into the periplasm, thereby allowing access to the Xcp (type II) secretory system for further export in Gramnegative organisms, or directly to the outside surface of the cell, as in M. tuberculosis. TAT-mediated secretion appreciably contributes to virulence in both animal and plant models of bacterial infection. Consequently, TAT function is an attractive target for small-molecular-weight compounds that alone or in conjunction with extant antimicrobial agents could become novel therapeutics. The TAT-transported hemolytic phospholipase C (PlcH) of P. aeruginosa and its multiple orthologs produced by the above pathogens can be detected by an accurate and reproducible colorimetric assay using a synthetic substrate that detects phospholipase C activity. Such an assay could be an effective indicator of TAT function. Using carefully constructed recombinant strains to precisely control the expression of PlcH, we developed a high-throughput screening (HTS) assay to evaluate, in duplicate, >80,000 small-molecular-weight compounds as possible TAT inhibitors. Based on additional TAT-related functional assays, purified PlcH protein inhibition experiments, and repeat experiments of the initial screening assay, 39 compounds were selected from the 122 initial hits. Finally, to evaluate candidate inhibitors for TAT specificity, we developed a TAT titration assay that determines whether inhibition of TAT-mediated secretion can be overcome by increasing the levels of TAT expression. The compounds N-phenyl maleimide and Bay 11-7082 appear to directly affect TAT function based on this approach.T he twin-arginine translocase (TAT) secretion system was first discovered as a Sec-independent mechanism by which proteins involved in photosynthesis are transported into the thylakoids of plant chloroplasts (10). Orthologs of plant TAT proteins were then identified in Escherichia coli and subsequently in other bacteria and archaea (58). However, no orthologs or functional analogs of TAT proteins have thus far been identified in animal cells.The TAT system in plants translocates proteins into the thylakoid compartment of chloroplasts, whereas in bacteria and archaea, it can export proteins out of the cytoplasm and across the cytoplasmic membrane. In Gram-negative organisms (e.g., E. coli, Pseudomonas aeruginosa), proteins secreted by the TAT system are most frequently terminally localized in the periplasm, but some (e.g., phospholipases C) can be further secreted by the Xcp (i.e., type II) system, thereby becoming extracellular (41,65,66). In Gram-positive bacteria (e.g., Bacillus spp., Mycobacterium spp., and Streptomycetes spp.) (26, 27, 36), TAT substrates typically remain cell associated upon secretion through the cytoplasmic membrane, but at least three proteins (i.e., agara...

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Structure analysis of the protein translocating channel TatA in membranes using a multi-construct approach

Cited by 42 publications

References 28 publications

Variable stoichiometry of the TatA component of the twin-arginine protein transport system observed by in vivo single-molecule imaging

Variable stoichiometry of the TatA component of the twin-arginine protein transport system observed by in vivo single-molecule imaging

Salt Sensitivity of Minimal Twin Arginine Translocases

Identification and Evaluation of Twin-Arginine Translocase Inhibitors

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