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Shearing and Enrichment of Extracellular Type IV Pili

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Bacterial Protein Secretion Systems

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1615))

Abstract

Pili are widespread among bacteria. Type IVa pili (T4aP) are associated with a variety of bacterial functions, including adhesion, motility, natural transformation, biofilm formation, and force-dependent signaling. In pathogenic bacteria, T4aP play a crucial role during infection and have been the subject of hundreds of studies. Methods for the isolation and purification of T4aP were first described in the 1970s. Purified pili have been used for studies of filament protein content, morphology, immunogenicity, post-translational modifications, and X-ray crystallography. We detail a tried-and-true method of isolating large amounts of native T4aP from bacterial surfaces. The method requires supplies and equipment that are available in most microbiology labs.

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Acknowledgements

We are grateful to Dr. Nicole Koropatkin for purifying grams of pili and optimizing this protocol in the process, and to Dr. Lisa Craig for many years of collegial interactions and helpful suggestions for this chapter.

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Authors and Affiliations

  1. Department of Bacteriology and Biophysics Program, University of Wisconsin-Madison, Madison, WI, 53706, USA

    Alba Katiria Gonzalez Rivera & Katrina T. Forest

Authors
  1. Alba Katiria Gonzalez Rivera
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  2. Katrina T. Forest
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Corresponding author

Correspondence to Katrina T. Forest .

Editor information

Editors and Affiliations

  1. Institut de Microbiologie de la Méditerranée, Aix-Marseille Univ - CNRS, Marseille, France

    Laure Journet

  2. Institut de Microbiologie de la Méditerranée, Aix-Marseille Univ - CNRS, Marseille, France

    Eric Cascales

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Gonzalez Rivera, A.K., Forest, K.T. (2017). Shearing and Enrichment of Extracellular Type IV Pili. In: Journet, L., Cascales, E. (eds) Bacterial Protein Secretion Systems. Methods in Molecular Biology, vol 1615. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7033-9_25

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  • DOI: https://doi.org/10.1007/978-1-4939-7033-9_25

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7031-5

  • Online ISBN: 978-1-4939-7033-9

  • eBook Packages: Springer Protocols

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