A potential late stage intermediate of twin-arginine dependent protein translocation in Escherichia coli

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dc.identifier.uri http://dx.doi.org/10.15488/5218
dc.identifier.uri https://www.repo.uni-hannover.de/handle/123456789/5265
dc.contributor.author Geise, Hendrik
dc.contributor.author Heidrich, Eyleen Sabine
dc.contributor.author Nikolin, Christoph Stefan
dc.contributor.author Mehner-Breitfeld, Denise
dc.contributor.author Brüser, Thomas
dc.date.accessioned 2019-08-26T07:56:06Z
dc.date.available 2019-08-26T07:56:06Z
dc.date.issued 2019
dc.identifier.citation Geise, H.; Heidrich, E.S.; Nikolin, C.S.; Mehner-Breitfeld, D.; Brüser, T.: A potential late stage intermediate of twin-arginine dependent protein translocation in Escherichia coli. In: Frontiers in Microbiology 10 (2019), 1482. DOI: https://doi.org/10.3389/fmicb.2019.01482
dc.description.abstract The twin-arginine translocation (Tat) system transports folded proteins across membranes of prokaryotes, plant plastids, and some mitochondria. According to blue-native polyacrylamide gel electrophoresis after solubilization with digitonin, distinct interactions between the components TatA, TatB, and TatC result in two major TatBC-containing complexes in Escherichia coli that can bind protein substrates. We now report the first detection of a TatABC complex that likely represents the state at which transport occurs. This complex was initially found when the photo cross-linking amino acid p-benzoyl-L-phenylalanine (Bpa) was introduced at position I50 on the periplasmic side of the first trans-membrane domain of TatC. Cross-linking of TatCI50Bpa resulted in TatC-TatC-cross-links, indicating a close proximity to neighboring TatC in the complex. However, the new complex was not caused by cross-links but rather by non-covalent side chain interactions, as it was also detectable without UV-cross-linking or with an I50Y exchange. The new complex did not contain any detectable substrate. It was slightly upshifted relative to previously reported substrate-containing TatABC complexes. In the absence of TatA, an inactive TatBCI50Bpa complex was formed of the size of wild-type substrate-containing TatABC complexes, suggesting that TatB occupies TatA-binding sites at TatCI50Bpa. When substrate binding was abolished by point mutations, this TatBCI50Bpa complex shifted analogously to active TatABCI50Bpa complexes, indicating that a defect substrate-binding site further enhances TatB association to TatA-binding sites. Only TatA could shift the complex with an intact substrate-binding site, which explains the TatA requirement for substrate transport by TatABC systems. eng
dc.language.iso eng
dc.publisher Lausanne : Frontiers Media S.A.
dc.relation.ispartofseries Frontiers in Microbiology 10 (2019)
dc.rights CC BY 4.0
dc.rights.uri https://creativecommons.org/licenses/by/4.0/
dc.subject Escherichia coli eng
dc.subject Membrane protein complexes eng
dc.subject Photo cross-linking eng
dc.subject Protein translocation eng
dc.subject Twin-arginine translocation eng
dc.subject digitonin eng
dc.subject n benzoylphenylalanine derivative eng
dc.subject protein translocase eng
dc.subject TATA binding protein eng
dc.subject TATA binding protein related factor eng
dc.subject affinity chromatography eng
dc.subject Article eng
dc.subject bacterial translocation eng
dc.subject binding site eng
dc.subject Escherichia coli eng
dc.subject gene expression eng
dc.subject gene mutation eng
dc.subject immunoblotting eng
dc.subject mutagenesis eng
dc.subject nonhuman eng
dc.subject phase contrast microscopy eng
dc.subject plasmid eng
dc.subject point mutation eng
dc.subject polyacrylamide gel electrophoresis eng
dc.subject protein binding eng
dc.subject protein cross linking eng
dc.subject protein expression eng
dc.subject size exclusion chromatography eng
dc.subject Western blotting eng
dc.subject.ddc 570 | Biowissenschaften, Biologie ger
dc.title A potential late stage intermediate of twin-arginine dependent protein translocation in Escherichia coli
dc.type article
dc.type Text
dc.relation.issn 1664-302X
dc.relation.doi https://doi.org/10.3389/fmicb.2019.01482
dc.bibliographicCitation.volume 10
dc.bibliographicCitation.firstPage 1482
dc.description.version publishedVersion
tib.accessRights frei zug�nglich


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