Abstract
The flagellar type III export apparatus utilizes ATP and proton motive force (PMF) across the cytoplasmic membrane as the energy sources and transports flagellar component proteins from the cytoplasm to the distal growing end of the growing structure to construct the bacterial flagellum beyond the cellular membranes. The flagellar type III export apparatus coordinates flagellar protein export with assembly by ordered export of substrates to parallel with their order of the assembly. The export apparatus is composed of a PMF-driven transmembrane export gate complex and a cytoplasmic ATPase complex. Since the ATPase complex is dispensable for flagellar protein export, PMF is the primary fuel for protein unfolding and translocation. Interestingly, the export gate complex can also use sodium motive force across the cytoplasmic membrane in addition to PMF when the ATPase complex does not work properly. Here, we describe experimental protocols, which have allowed us to identify the export substrate class and the primary fuel of the flagellar type III protein export apparatus in Salmonella enterica serovar Typhimurium.
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Acknowledgments
This research has been supported in part by JSPS KAKENHI Grant Numbers JP26293097 (to T.M.) and JP25000013 (to K.N.) and MEXT KAKENHI Grant Numbers JP25121718 and JP15H01640 (to T.M.).
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Minamino, T., Kinoshita, M., Namba, K. (2017). Fuel of the Bacterial Flagellar Type III Protein Export Apparatus. In: Minamino, T., Namba, K. (eds) The Bacterial Flagellum. Methods in Molecular Biology, vol 1593. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6927-2_1
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DOI: https://doi.org/10.1007/978-1-4939-6927-2_1
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