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Purification of LPS from Leptospira

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Leptospira spp.

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2134))

Abstract

Leptospira species are one of the few spirochetes to possess a lipopolysaccharide (LPS) embedded in their outer membrane. Two protocols are currently available to extract and/or purify the leptospiral lipopolysaccharides: the rapid proteinase K method and the classical hot water/phenol extraction. The first method allows to get a quick overview of the LPS O antigen structure, whereas the second method is fitted to study the immunological properties of the leptospiral LPS. These two methods will be detailed in this chapter. Methodologies to assess the quality of the purification, such as the modified silver staining coloration, will also be reviewed. Both advantages and limitations of the different analyses will be described.

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References

  1. Murray GL, Srikram A, Henry R, Hartskeerl RA, Sermswan RW, Adler B (2010) Mutations affecting Leptospira interrogans lipopolysaccharide attenuate virulence. Mol Microbiol 78(3):701–709

    Article  CAS  Google Scholar 

  2. Que-Gewirth NL, Ribeiro AA, Kalb SR, Cotter RJ, Bulach DM, Adler B et al (2004) A methylated phosphate group and four amide-linked acyl chains in Leptospira interrogans lipid A. The membrane anchor of an unusual lipopolysaccharide that activates TLR2. J Biol Chem 279(24):25420–25429

    Article  CAS  Google Scholar 

  3. Werts C (2010) Leptospirosis: a Toll road from B lymphocytes. Chang Gung Med J 33(6):591–601

    PubMed  Google Scholar 

  4. Nahori MA, Fournie-Amazouz E, Que-Gewirth NS, Balloy V, Chignard M, Raetz CR et al (2005) Differential TLR recognition of leptospiral lipid A and lipopolysaccharide in murine and human cells. J Immunol 175(9):6022–6031

    Article  CAS  Google Scholar 

  5. Werts C, Tapping RI, Mathison JC, Chuang TH, Kravchenko V, Saint Girons I et al (2001) Leptospiral lipopolysaccharide activates cells through a TLR2-dependent mechanism. Nat Immunol 2(4):346–352

    Article  CAS  Google Scholar 

  6. Westphal O, Lüderitz O, Bister F (1952) Über die Extraktion von Bakterien mit Phenol/Wasser. Z Naturforsch 7(3):148–155

    Article  Google Scholar 

  7. Eshghi A, Henderson J, Trent MS, Picardeau M (2015) Leptospira interrogans lpxD homologue is required for thermal acclimatization and virulence. Infect Immun 83(11):4314–4321

    Article  CAS  Google Scholar 

  8. Tsai CM, Frasch CE (1982) A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal Biochem 119(1):115–119

    Article  CAS  Google Scholar 

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Correspondence to Catherine Werts .

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Bonhomme, D., Werts, C. (2020). Purification of LPS from Leptospira. In: Koizumi, N., Picardeau, M. (eds) Leptospira spp.. Methods in Molecular Biology, vol 2134. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0459-5_6

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  • DOI: https://doi.org/10.1007/978-1-0716-0459-5_6

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0458-8

  • Online ISBN: 978-1-0716-0459-5

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