Prokaryotic organisms possess a specialized protein translocase in their cytoplasmic membranes that catalyzes the export of folded preproteins. Substrates for this pathway are distinguished by a twin-arginine consensus motif in their signal peptides (twin-arginine translocation [Tat] pathway). We have compiled detailed protocols for the preparation and operation of a cell-free system by which the bacterial Tat pathway can be fully reproduced in vitro. This system has proven useful and is being further exploited for the study of precursor–translocase interactions, assembly of the translocase, and the mechanism of transmembrane passage.
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Acknowledgments
This work was supported by grant LSHG-CT-2004-05257 of the European Union and grants from the Deutsche Forschungsgemeinschaft (Sonderforschungsbereich 388 and Graduiertenkolleg 434).
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Moser, M., Panahandeh, S., Holzapfel, E., Müller, M. (2007). In Vitro Analysis of the Bacterial Twin-Arginine-Dependent Protein Export. In: van der Giezen, M. (eds) Protein Targeting Protocols. Methods in Molecular Biology™, vol 390. Humana Press. https://doi.org/10.1007/978-1-59745-466-7_5
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DOI: https://doi.org/10.1007/978-1-59745-466-7_5
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