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Characterization of the type III export signal of the flagellar hook scaffolding protein FlgD of Escherichia coli

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Abstract

Transport of flagellar structural proteins beyond the cytoplasmic membrane is accomplished by a type III secretory pathway [flagellar type III secretion system (fTTSS)]. The mechanism of substrate recognition by the fTTSS is still enigmatic. Using the hook scaffolding protein FlgD of Escherichia coli as a model substrate, it is demonstrated that the export signal is contained within the N-terminal 71 amino acids of FlgD. Analysis of frame-shift mutations and alterations of the nucleotide sequence suggest a proteinaceous nature of the signal. Furthermore, the physicochemical properties of the first about eight amino acids are crucial for export.

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Abbreviations

BCIP:

5-Bromo-4-chloro-3-indolyl phosphate

ICDH:

Isocitrate dehydrogenase

IPTG:

Isopropyl 1-thio-β-d-galactopyranoside

LB:

Luria-Bertani broth

PAGE:

Polyacrylamide gel electrophoresis

pNPP:

p-Nitrophenyl phosphate

SDS:

Sodium dodecyl sulfate

TTSS:

Type III secretion system

fTTSS:

Flagellar TTSS

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Acknowledgments

We thank Silvia Wagner for technical assistance. This work was financially supported by the Deutsche Forschungsgemeinschaft (Ju333/2).

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Author notes

  1. Petra Pöplau, Carolin Möckel & Heinrich Jung

    Present address: Department Biology I, Microbiology, LMU Munich, Maria-Ward-Strasse 1a, 80638, Munich, Germany

  2. Maike Rochón

    Present address: Gesellschaft für Biotechnologische Forschung mbH, Mascheroder Weg 1, 38124, Braunschweig, Germany

Authors and Affiliations

  1. Fachbereich Biologie/Chemie, Abteilung Mikrobiologie, Universität Osnabrück, Barbarastrasse 11, 49069, Osnabrück, Germany

    Corinna Weber-Sparenberg, Petra Pöplau, Heiner Brookman, Maike Rochón, Monika Nietschke & Heinrich Jung

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  1. Corinna Weber-Sparenberg
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  2. Petra Pöplau
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  3. Heiner Brookman
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Correspondence to Heinrich Jung.

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Weber-Sparenberg, C., Pöplau, P., Brookman, H. et al. Characterization of the type III export signal of the flagellar hook scaffolding protein FlgD of Escherichia coli . Arch Microbiol 186, 307–316 (2006). https://doi.org/10.1007/s00203-006-0146-0

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  • DOI: https://doi.org/10.1007/s00203-006-0146-0

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