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Characterization of Type Three Secretion System Translocator Interactions with Phospholipid Membranes

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Type 3 Secretion Systems

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1531))

Abstract

In vitro characterization of type III secretion system (T3SS) translocator proteins has proven challenging due to complex purification schemes and their hydrophobic nature that often requires detergents to provide protein solubility and stability. Here, we provide experimental details for several techniques that overcome these hurdles, allowing for the direct characterization of the Shigella translocator protein IpaB with respect to phospholipid membrane interaction. The techniques specifically discussed in this chapter include membrane interaction/liposome flotation, liposome sensitive fluorescence quenching, and protein-mediated liposome disruption assays. These assays have provided valuable insight into the role of IpaB in T3SS-mediated phospholipid membrane interactions by Shigella and should readily extend to other members of this important class of proteins.

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References

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Author information

Authors and Affiliations

  1. Kansas Department of Health and Environment Laboratories, Topeka, KS, 66620, USA

    Philip R. Adam

  2. Higuchi Biosciences Center, University of Kansas, Lawrence, KS, 66047, USA

    Michael L. Barta

  3. Department of Chemistry and Biochemistry, Utah State University, 300 Old Main Hill, Logan, UT, 84322, USA

    Nicholas E. Dickenson

Authors
  1. Philip R. Adam
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  2. Michael L. Barta
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  3. Nicholas E. Dickenson
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Corresponding author

Correspondence to Nicholas E. Dickenson .

Editor information

Editors and Affiliations

  1. Department of Biomedical Sciences, School of Medicine and Health Sciences University of North Dakota, Grand Forks, North Dakota, USA

    Matthew L. Nilles

  2. Department of Biomedical Sciences, School of Medicine and Health Sciences University of North Dakota, Grand Forks, North Dakota, USA

    Danielle L. Jessen Condry

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© 2017 Springer Science+Business Media New York

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Adam, P.R., Barta, M.L., Dickenson, N.E. (2017). Characterization of Type Three Secretion System Translocator Interactions with Phospholipid Membranes. In: Nilles, M., Condry, D. (eds) Type 3 Secretion Systems. Methods in Molecular Biology, vol 1531. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6649-3_7

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  • DOI: https://doi.org/10.1007/978-1-4939-6649-3_7

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6647-9

  • Online ISBN: 978-1-4939-6649-3

  • eBook Packages: Springer Protocols

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