Chapter title |
A Coculture-Based Approach for Screening Campaigns Aimed at Identifying Novel Pseudomonas aeruginosa Quorum Sensing Inhibitors
|
---|---|
Chapter number | 22 |
Book title |
Quorum Sensing
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7309-5_22 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7308-8, 978-1-4939-7309-5
|
Authors |
Giordano Rampioni, Giulia Giallonardi, Francesca D’Angelo, Livia Leoni |
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. |
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