Abstract
The Type IV pili are displayed peritrichously on the surfaces of Neisseria gonorrhoeae cells. Here we present protocols for isolating and purifying Type IV pili and dissociating them into PilE pilin subunits. Pilus filaments are isolated from the bacterial cell surface by mechanical shearing and purified by differential precipitation and centrifugation. PilE subunits are extracted by treating the purified pili with detergent to disrupt the hydrophobic interactions holding them together in the filaments. Purified pili and pilin subunits can be used for structural, biophysical, or biochemical characterization and as antigens for antibody production.
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Acknowledgments
This work was supported by the Canadian Institutes of Health Research (MOP125959). We thank Katrina Forest, University of Wisconsin-Madison, Wisconsin, the USA, for helpful discussions.
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Craig, L., Altindal, T. (2019). Purification of Type IV Pili and Pilin Subunits. In: Christodoulides, M. (eds) Neisseria gonorrhoeae. Methods in Molecular Biology, vol 1997. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9496-0_7
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DOI: https://doi.org/10.1007/978-1-4939-9496-0_7
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Publisher Name: Humana, New York, NY
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