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Quorum Sensing pp 287-296 | Cite as

A Coculture-Based Approach for Screening Campaigns Aimed at Identifying Novel Pseudomonas aeruginosa Quorum Sensing Inhibitors

  • Giordano Rampioni
  • Giulia Giallonardi
  • Francesca D’Angelo
  • Livia Leoni
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1673)

Abstract

Quorum sensing (QS) is recognized as a promising target for the identification of anti-virulence drugs hampering Pseudomonas aeruginosa adaptability to the host environment and pathogenicity. Consequently, a number of studies in the last decade focused on the identification of small molecules or proteins with anti-QS activity, mainly targeting the las QS system, which is based on N-3-oxododecanoyl-homoserine lactone (3OC12-HSL) as signal molecule. Different experimental approaches have been successfully used to identify QS blockers interfering with the activity/stability of the 3OC12-HSL receptor LasR, with the functionality of the 3OC12-HSL synthase LasI, or with the stability/bioavailability of the 3OC12-HSL signal molecule itself.

Here we describe the use of a high-throughput screening system for the identification of novel las QS inhibitors based on the cocultivation of P. aeruginosa wild type and the P. aeruginosa-derived biosensor strain PA14-R3, in which light emission relies on the ability of the wild type strain to synthesize 3OC12-HSL and of the biosensor strain to perceive this signal molecule. With respect to other screening systems, this method has the advantage of being cost-effective and allowing the identification of compounds targeting, besides 3OC12-HSL reception, any cellular process critical for the functionality of the las QS system, including 3OC12-HSL synthesis and secretion.

Key words

Quorum sensing inhibitors Screening Whole-cell biosensors Anti-virulence drugs Niclosamide Pseudomonas aeruginosa lasR lasI 

Notes

Acknowledgments

This work was supported by the Italian Ministry for University and Research (RBFR10LHD1 to G.R.), and by the Italian Cystic Fibrosis Research Foundation (FFC 10/2013 to L.L.).

We wish to thanks all the colleagues contributing to the development of PA14-R3 and to its application in screening campaigns: Francesco Imperi, Francesco Massai, Francesca Longo, Cejoice Ramachandran Pillai, Elisabetta Zennaro, and Paolo Visca.

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Copyright information

© Springer Science+Business Media LLC 2018

Authors and Affiliations

  • Giordano Rampioni
    • 1
  • Giulia Giallonardi
    • 1
  • Francesca D’Angelo
    • 1
  • Livia Leoni
    • 1
  1. 1.Department of ScienceUniversity Roma TreRomeItaly

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