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Quantitative and Qualitative Preparations of Bacterial Outer Membrane Vesicles

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Bacterial Cell Surfaces

Part of the book series: Methods in Molecular Biology ((MIMB,volume 966))

Abstract

Gram-negative bacterial outer membrane vesicle production and function have been studied using a variety of quantitative and qualitative methods. These types of analyses can be hampered by the use of impure vesicle preparations. Here we describe a set of techniques that are useful for the quantitative analysis of vesicle production and for preparative yields of highly purified vesicles for studies of vesicle function or composition. Procedures and advice are also included for the purification of vesicles from encapsulated and low-yield strains.

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References

  1. Kulp A, Kuehn MJ (2010) Biological functions and biogenesis of secreted bacterial outer membrane vesicles. Annu Rev Microbiol 64:163–184

    Article  PubMed  CAS  Google Scholar 

  2. Kuehn MJ, Kesty NC (2005) Bacterial outer membrane vesicles and the host-pathogen interaction. Genes Dev 19(22):2645–2655

    Article  PubMed  CAS  Google Scholar 

  3. Lee EY, Choi DS, Kim KP, Gho YS (2008) Proteomics in gram-negative bacterial outer membrane vesicles. Mass Spectrom Rev 27(6):535–555

    Article  PubMed  CAS  Google Scholar 

  4. Beveridge TJ (1999) Structures of gram-negative cell walls and their derived membrane vesicles. J Bacteriol 181(16):4725–4733

    PubMed  CAS  Google Scholar 

  5. Bishop DG, Work E (1965) An extracellular glycolipid produced by Escherichia coli grown under lysine-limiting conditions. Biochem J 96(2):567–576

    PubMed  CAS  Google Scholar 

  6. McBroom AJ, Johnson AP, Vemulapalli S, Kuehn MJ (2006) Outer membrane vesicle production by Escherichia coli is independent of membrane instability. J Bacteriol 188(15):5385–5392

    Article  PubMed  CAS  Google Scholar 

  7. Ciofu O, Beveridge TJ, Kadurugamuwa J, Walther-Rasmussen J, Høiby N (2000) Chromosomal β-lactamase is packaged into membrane vesicles and secreted from Pseudomonas aeruginosa. J Antimicrob Chemother 45(1):9–13

    Article  PubMed  CAS  Google Scholar 

  8. Schooling SR, Beveridge TJ (2006) Membrane vesicles: an overlooked component of the matrices of biofilms. J Bacteriol 188(16):5945–5957

    Article  PubMed  CAS  Google Scholar 

  9. Hellman J, Loiselle PM, Zanzot EM, Allaire JE, Tehan MM, Boyle LA, Kurnick JT, Warren HS (2000) Release of gram-negative outer-membrane proteins into human serum and septic rat blood and their interactions with immunoglobulin in antiserum to Escherichia coli J5. J Infect Dis 181(3):1034–1043

    Article  PubMed  CAS  Google Scholar 

  10. McBroom AJ, Kuehn MJ (2007) Release of outer membrane vesicles by Gram-negative bacteria is a novel envelope stress response. Mol Microbiol 63(2):545–558

    Article  PubMed  CAS  Google Scholar 

  11. Chutkan H, Kuehn MJ (2011) Context-dependent activation kinetics elicited by soluble versus outer membrane vesicle-associated heat-labile enterotoxin. Infect Immun 79(9):3760–3769

    Article  PubMed  CAS  Google Scholar 

  12. Ellis TN, Leiman SA, Kuehn MJ (2010) Naturally produced outer membrane vesicles from Pseudomonas aeruginosa elicit a potent innate immune response via combined sensing of both lipopolysaccharide and protein components. Infect Immun 78(9):3822–3831

    Article  PubMed  CAS  Google Scholar 

  13. Bauman SJ, Kuehn MJ (2006) Purification of outer membrane vesicles from Pseudomonas aeruginosa and their activation of an IL-8 response. Microbes Infect 8(9–10):2400–2408

    Article  PubMed  CAS  Google Scholar 

  14. Bauman SJ, Kuehn MJ (2009) Pseudomonas aeruginosa vesicles associate with and are internalized by human lung epithelial cells. BMC Microbiol 9:26

    Article  PubMed  Google Scholar 

  15. Shoemaker DR, Saunders NB, Brandt BL, Moran EE, LaClair AD, Zollinger WD (2005) Intranasal delivery of group B meningococcal native outer membrane vesicle vaccine induces local mucosal and serum bactericidal antibody responses in rabbits. Infect Immun 73(8):5031–5038

    Article  PubMed  CAS  Google Scholar 

  16. Manning AJ, Kuehn MJ (2011) Contribution of bacterial outer membrane vesicles to innate bacterial defense. BMC Microbiol 11:258

    Article  PubMed  CAS  Google Scholar 

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Acknowledgements

This work was supported by NIH grants R01AI079068, R01AI064464, and R01GM099471-01. We thank Harold Meekel (University of North Carolina-Chapel Hill) for assistance with electron microscopy.

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Authors and Affiliations

  1. Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, NC, USA

    Halima Chutkan & Ian MacDonald

  2. Department of Biochemistry, Duke University Medical Center, Durham, NC, USA

    Andrew Manning & Meta J. Kuehn

Authors
  1. Halima Chutkan
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  2. Ian MacDonald
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  3. Andrew Manning
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  4. Meta J. Kuehn
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Corresponding author

Correspondence to Meta J. Kuehn .

Editor information

Editors and Affiliations

  1. , Dept. of Biology and Biochemistry, University of Houston, Calhoun Road 4800, Houston, 77004, Texas, USA

    Anne H. Delcour

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Chutkan, H., MacDonald, I., Manning, A., Kuehn, M.J. (2013). Quantitative and Qualitative Preparations of Bacterial Outer Membrane Vesicles. In: Delcour, A. (eds) Bacterial Cell Surfaces. Methods in Molecular Biology, vol 966. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-245-2_16

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  • DOI: https://doi.org/10.1007/978-1-62703-245-2_16

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-244-5

  • Online ISBN: 978-1-62703-245-2

  • eBook Packages: Springer Protocols

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