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Quorum Sensing pp 291-298 | Cite as

High-Performance Liquid Chromatography Analysis of N-Acyl Homoserine Lactone Hydrolysis by Paraoxonases

  • John F. Teiber
  • Dragomir I. Draganov
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 692)

Abstract

Mammalian paraoxonases (PONs) are a unique, highly conserved family of calcium-dependent esterases consisting of PON1, PON2, and PON3. The PONs can hydrolyze the lactone ring of a range of N-acyl-l-homoserine lactone (AHL) quorum sensing signaling molecules, rendering them inactive. This chapter describes a method that utilizes high-performance liquid chromatography analysis with UV detection for determining the rate of AHL hydrolysis in cell lysates, tissue homogenates, serum, and with purified proteins. Also described are the techniques used to prepare cell culture lysates and tissue homogenates for analysis and the use of class-specific enzyme inhibitors to determine the contribution of PONs to AHL hydrolysis in the samples.

Key words

Acyl Homoserine Lactone High-performance liquid chromatography Lactonase Paraoxonases Quorum sensing 

References

  1. 1.
    Draganov DI, Teiber JF, Speelman A, Osawa Y, Sunahara R, La Du BN. (2005) Human paraoxonases (PON1, PON2, and PON3) are lactonases with overlapping and distinct substrate specificities. J Lipid Res; 46(6):1239–1247.PubMedCrossRefGoogle Scholar
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    Draganov D, Teiber J. (2008) PONs’ natural substrates-the key for their physiological roles. In: Mackness M, Aviram M, Parah G, editors. Protein and Cell Regulation, Volume 6: The Paraoxonases: Their Role in Disease Develop ment and Xenobiotic Metabolism. Dordrecht: Springer: 297–306.Google Scholar
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    Stoltz DA, Ozer EA, Ng CJ et al. (2007) Paraoxonase-2 deficiency enhances Pseudo monas aeruginosa quorum sensing in murine tracheal epithelia. Am J Physiol Lung Cell Mol Physiol; 292(4):L852–L860.PubMedCrossRefGoogle Scholar
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    Teiber JF, Horke S, Haines DC et al. (2008) Dominant role of paraoxonases in inactivation of the Pseudomonas aeruginosa quorum-sensing signal N-(3-oxododecanoyl)-L-homoserine lactone. Infect Immun; 76(6):2512–2519.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • John F. Teiber
    • 1
  • Dragomir I. Draganov
    • 1
  1. 1.Division of Epidemiology, Department of Internal MedicineThe University of Texas Southwestern Medical CenterDallasUSA

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