Abstract
Many motile bacteria employ the flagellar type III secretion system (fT3SS) to build the flagellum on the cell surface. The fT3SS consists of a transmembrane export gate complex, which acts as a proton/protein antiporter that couples proton flow with flagellar protein export, and a cytoplasmic ATPase ring complex, which works as an activator of the export gate complex. Three transmembrane proteins, FliP, FliQ, and FliR, form a core structure of the export gate complex, and this core complex serves as a polypeptide channel that allows flagellar structural subunits to be translocated across the cytoplasmic membrane. Here, we describe the methods for overproduction, solubilization, and purification of the Salmonella FliP/FliQ/FliR complex.
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Acknowledgments
This work was supported in part by JSPS KAKENHI Grant Numbers JP20K15749 and JP22K06162 (to M.K.) and JP19H03182, JP22H02573, and JP22K19274 (to T.M.) and MEXT KAKENHI Grant Number JP20H05532 and JP22H04844 (to T.M.). This work has also been supported by Platform Project for Supporting Drug Discovery and Life Science Research (BINDS) from AMED under Grant Number JP19am0101117 to K.N., by the Cyclic Innovation for Clinical Empowerment (CiCLE) from AMED under Grant Number JP17pc0101020 to K.N. and by JEOL YOKOGUSHI Research Alliance Laboratories of Osaka University to K.N.
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Kinoshita, M., Namba, K., Minamino, T. (2023). Purification of the Transmembrane Polypeptide Channel Complex of the Salmonella Flagellar Type III Secretion System. In: Minamino, T., Miyata, M., Namba, K. (eds) Bacterial and Archaeal Motility. Methods in Molecular Biology, vol 2646. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3060-0_1
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DOI: https://doi.org/10.1007/978-1-0716-3060-0_1
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