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 |
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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 |
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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 |
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dc.publisher |
Lausanne : Frontiers Media S.A. |
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dc.relation.ispartofseries |
Frontiers in Microbiology 10 (2019) |
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dc.rights |
CC BY 4.0 Unported |
|
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 |
eng |
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 |
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dc.description.version |
publishedVersion |
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tib.accessRights |
frei zug�nglich |
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