Purification of Native Flagellin

  • María José González
  • Victoria Iribarnegaray
  • Pablo Zunino
  • Paola Scavone
Part of the Methods in Molecular Biology book series (MIMB, volume 2021)


Flagella are effective organelles of locomotion and one of several virulence factors in Proteus mirabilis. To study their properties and role in virulence, we describe a protocol to extract and purify the native flagellin of P. mirabilis. Purified flagellin can be visualized by SDS-PAGE or immunoblot and is suitable for downstream applications such as immunization.

Key words

Proteus mirabilis Native flagellin Flagellum SDS-PAGE Western blot 


  1. 1.
    Sourjik V, Wingreen NS (2012) Responding to chemical gradients: bacterial chemotaxis. Curr Opinion Cell Biol 24(2):262–268CrossRefGoogle Scholar
  2. 2.
    Berg HC (2003) The rotary motor of bacterial flagella. Annu Rev Biochem 72:19–54CrossRefGoogle Scholar
  3. 3.
    Evans LD, Poulter S, Terentjev EM, Hughes C, Fraser GM (2013) A chain mechanism for flagellum growth. Nature 504(7479):287–290CrossRefGoogle Scholar
  4. 4.
    Rossez Y, Wolfson EB, Holmes A, Gally DL, Holden NJ (2015) Bacterial flagella: twist and stick, or dodge across the kingdoms. PLoS Path 11(1):e1004483CrossRefGoogle Scholar
  5. 5.
    Chevance FF, Hughes KT (2008) Coordinating assembly of a bacterial macromolecular machine. Nature Rev Microbiol 6(6):455–465CrossRefGoogle Scholar
  6. 6.
    Mizel SB, Bates JT (2010) Flagellin as an adjuvant: cellular mechanisms and potential. J Immunol 185(10):5677–5682CrossRefGoogle Scholar
  7. 7.
    Macnab RM (2003) How bacteria assemble flagella. Annu Rev Microbiol 57:77–100CrossRefGoogle Scholar
  8. 8.
    Rather PN (2005) Swarmer cell differentiation in Proteus mirabilis. Environ Microbiol 7(8):1065–1073CrossRefGoogle Scholar
  9. 9.
    Schaffer JN, Pearson MM (2015) Proteus mirabilis and urinary tract infections. Microbiology Spectrum 3(5)Google Scholar
  10. 10.
    Pearson MM, Yep A, Smith SN, Mobley HLT (2011) Transcriptome of Proteus mirabilis in the murine urinary tract: virulence and nitrogen assimilation gene expression. Infect Immun 79(7):2619–2631CrossRefGoogle Scholar
  11. 11.
    Legnani-Fajardo C, Zunino P, Algorta G, Laborde HF (1991) Antigenic and immunogenic activity of flagella and fimbriae preparations from uropathogenic Proteus mirabilis. Canadian J Microbiol 37(4):325–328CrossRefGoogle Scholar
  12. 12.
    Scavone P, Umpiérrez A, Rial A, Chabalgoity JA, Zunino P (2014) Native flagellin does not protect mice against an experimental Proteus mirabilis ascending urinary tract infection and neutralizes the protective effect of MrpA fimbrial protein. Antonie van Leeuwenhoek 105(6):1139–1148CrossRefGoogle Scholar
  13. 13.
    Umpiérrez A, Scavone P, Romanin D, Marqués JM, Chabalgoity JA, Rumbo M, Zunino P (2013) Innate immune responses to Proteus mirabilis flagellin in the urinary tract. Microb Infect 15(10–11):688–696CrossRefGoogle Scholar
  14. 14.
    Murphy CA, Belas R (1999) Genomic rearrangements in the flagellin genes of Proteus mirabilis. Mol Microbiol 31:679–690CrossRefGoogle Scholar
  15. 15.
    Legnani-Fajardo C, Zunino P, Piccini C, Allen A, Maskell D (1996) Defined mutants of Proteus mirabilis lacking flagella cause ascending urinary tract infection in mice. Microb Pathog 21:395–405CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • María José González
    • 1
  • Victoria Iribarnegaray
    • 1
  • Pablo Zunino
    • 1
  • Paola Scavone
    • 1
  1. 1.Departamento de MicrobiologíaInstituto de Investigaciones Biológicas Clemente EstableMontevideoUruguay

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