Table Of ContentUse of Whole Cell Bioassays for Screening Quorum Signaling, Quorum-Interference, and Biofilm Dispersion.- Detection of 2-Alkyl-4-Quinolones Using Biosensors.- "Hot Stuff": The Many Uses of a Radiolabel Assay in Detecting Acyl-Homoserine Lactone Quorum Sensing Signals.- Liquid Chromatography/Mass Spectrometry (LC/MS) for the Detection and Quantification of N -Acyl-L-Homoserine Lactones (AHLs) and 4-Hydroxy-2-Alkylquinolines (HAQs).- Detection of the Bacterial Quorum Sensing Signaling Molecules N -Acyl Homoserine Lactones (HSL) and N -Acyl-Homoserine (HS) with an Enzyme-Linked Immunosorbent Assay (ELISA) and via Ultrahigh Performance Liquid Chromatography Coupled to Mass Spectrometry (UHPLC-MS).- Biosensors for the Detection and Quantification of AI-2 Class Quorum Sensing Compounds.- Detection of Agr-Type Autoinducing Peptides Produced by Staphylococcus aureus.- Ultra Performance Liquid Chromatography/Mass Spectrometryfor the Detection and Quantification of Diffusible Signal Factor (DSF)-Family Quorum Sensing Signals.- Rapid Electrochemical Detection of Pseudomonas aeruginosa Signaling Molecules by Boron-Doped Diamond Electrode.- Detection and Quantification of Butyrolactones from Streptomyces.- Fluorescence Quenching Studies of γ-Butyrolactone Binding Protein (CprB) from Streptomyces coelicolor A3(2).- Methods to Study Solo/Orphan Quorum Sensing Receptors.- Enzymatic Assays to Investigate Acyl-Homoserine Lactone Autoinducer Synthases.- Global Expression Analysis of Quorum Sensing-Controlled Genes by RNAseq.- Identification of AHL- and BDSF-Controlled Proteins in Burkholderia cenocepacia by Proteomics.- Imaging N -Acyl Homoserine Lactone Quorum Sensing In Vivo.- Assessing Pseudomonas aeruginosa Autoinducer Effects on Mammalian Epithelial Cells.- Animal Models for Pseudomonas aeruginosa Quorum Sensing Studies.- Methods to Study Quorum Sensing-Dependent Virulence and Movement of Phytopathogens In Planta.- Differential Equations Models to Study Quorum Sensing.- Qualitative and Quantitative Determination of Quorum Sensing Inhibition In Vitro.- A Co-Culture-Based Approach for Screening Campaigns Aimed at Identifying Novel Pseudomonas aeruginosa Quorum Sensing Inhibitors.- A Culture-Dependent Method for the Identification of Quorum Quenching Enzymes of Microbial Origin.- Directed Evolution of Quorum Quenching Enzymes: A Method for the Construction of a Directed Evolution Platform and Characterization of a Quorum-Quenching Lactonase from Geobacillus kaustophilus.- Generation of High-Sensitivity Monoclonal Antibodies Specific for Homoserine Lactones.- Identification of AI-2 Quorum Sensing Inhibitors in Vibrio harveyi through Structure-Based Virtual Screening.- Identification of Staphylococcal Quorum Sensing Inhibitors byQuantification of õ-Hemolysin with High Performance Liquid Chromatography.
SynopsisThis detailed volume provides scientists interested in quorum sensing with a broad spectrum of methods and protocols useful for studying bacterial communication processes at the chemico-physical, molecular, and physiological level. Divided into three sections, the content covers detection and quantification of quorum sensing signal molecules, methods for the studying of quorum sensing at the molecular, physiological, and population level, as well as identification and characterization of anti-quorum sensing agents. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Quorum Sensing: Methods and Protocols serves as a comprehensive guide to the most important methodology currently available in the field., Part I: Detection and Quantification of Quorum Sensing Signal Molecules 1. Use of Whole Cell Bioassays for Screening Quorum Signaling, Quorum-Interference, and Biofilm Dispersion Starla G. Thornhill and Robert J.C. McLean 2. Detection of 2-Alkyl-4-Quinolones Using Biosensors Matthew P. Fletcher, Stephen P. Diggle, Miguel Cámara, and Paul Williams 3. "Hot Stuff" The Many Uses of a Radiolabel Assay in Detecting Acyl-Homoserine Lactone Quorum Sensing Signals Amy L. Schaefer, Caroline S. Harwood, and E. Peter Greenberg 4. Liquid Chromatography/Mass Spectrometry (LC/MS) for the Detection and Quantification of -Acyl-L-Homoserine Lactones (AHLs) and 4-Hydroxy-2-Alkylquinolines (HAQs) François Lépine, Sylvain Milot, Marie-Christine Groleau, and Eric Déziel 5. Detection of the Bacterial Quorum Sensing Signaling Molecules N -Acyl Homoserine Lactones (HSL) and N -Acyl-Homoserine (HS) with an Enzyme-Linked Immunosorbent Assay (ELISA) and via Ultrahigh Performance Liquid Chromatography Coupled to Mass Spectrometry (UHPLC-MS) Michael Rothballer, Jenny Uhl, Josie Kunze, Philippe Schmitt-Kopplin, and Anton Hartmann 6. Biosensors for the Detection and Quantification of AI-2 Class Quorum Sensing Compounds Sathish Rajamani and Richard Sayre 7. Detection of Agr-Type Autoinducing Peptides Produced by Staphylococcus aureus Ewan J. Murray and Paul Williams 8. Ultra Performance Liquid Chromatography/Mass Spectrometry for the Detection and Quantification of Diffusible Signal Factor (DSF)-Family Quorum Sensing Signals Lian Zhou, Shuang Sun, Wei Zhang, and Ya-Wen He 9. Rapid Electrochemical Detection of Pseudomonas aeruginosa Signaling Molecules by Boron-Doped Diamond Electrode Alyah Buzid, John H.T. Luong, F. Jerry Reen, Fergal O'Gara, Jeremy D. Glennon, and Gerard P. McGlacken 10. Detection and Quantification of Butyrolactones from Streptomyces Marc Biarnes-Carrera, Rainer Breitling, and Eriko Takano Part II: Methods for the Studying of Quorum Sensing at the Molecular, Physiological, and Population Level 11. Fluorescence Quenching Studies of -Butyrolactone Binding Protein (CprB) from Streptomyces coelicolor A3(2) Jessy Mariam and Ruchi Anand 12. Methods to Study Solo/Orphan Quorum Sensing Receptors Vittorio Venturi, Sujatha Subramoni, Anice Sabag-Daigle, and Brian M.M. Ahmer 13. Enzymatic Assays to Investigate Acyl-Homoserine Lactone Autoinducer Synthases Daniel Shin and Rajesh Nagarajan 14. Global Expr