Abstract
Bacteria use chemical molecules called autoinducers as votes to poll their numerical strength in a colony. This polling mechanism, commonly referred to as quorum sensing, enables bacteria to build a social network and provide a collective response for fighting off common threats. In Gram-negative bacteria, AHL synthases synthesize acyl-homoserine lactone (AHL) autoinducers to turn on the expression of several virulent genes including biofilm formation, protease secretion, and toxin production. Therefore, inhibiting AHL signal synthase would limit quorum sensing and virulence. In this chapter, we describe four enzymatic methods that could be adopted to investigate a broad array of AHL synthases. The enzymatic assays described here should accelerate our mechanistic understanding of quorum-sensing signal synthesis that could pave the way for discovery of potent antivirulence compounds.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Bassler BL, Losick R (2006) Bacterially speaking. Cell 125:237–246
Schuster M, Lostroh CP, Ogi T, Greenberg EP (2003) Identification, timing, and signal specificity of Pseudomonas aeruginosa quorumcontrolled genes: a transcriptome analysis. J Bacteriol 185:2066–2079
Minogue TD, Wehland-von Trebra M, Bernhard F, von Bodman SB (2002) The autoregulatory role of EsaR, a quorum sensing regulator in Pantoea stewartii spp. stewartii: evidence for a repressor function. Mol Microbiol 44:1625–1635
De Kievit TR, Iglewski BH (2000) Bacterial quorum sensing in pathogenic relationships. Infect Immun 68:4839–4849
Chang Y-C, Krishnan T, Wang H, Chen Y, Yin W-F, Chong Y-M et al (2014) Non-antibiotic quorum sensing inhibitors acting against N-acyl homoserine lactone synthases as druggable target. Sci Rep 4:7245
Shin D, Frane ND, Brecht RM, Keeler J, Nagarajan R (2015) A comparative analysis of acyl-homoserine lactone synthase assays. ChemBioChem 16:2651–2659
Val DL, Cronan JE Jr (1998) In vivo evidence that S-adenosylmethionine and fatty acid synthesis intermediates are the substrates for the LuxI family of autoinducer synthases. J Bacteriol 180:2644–2651
Parsek MR, Val DL, Hanzelka BL, Cronan JE Jr, Greenberg EP (1999) Acyl homoserine-lactone quorum-sensing signal generation. Proc Natl Acad Sci U S A 96:4360–4365
Lindemann A, Pessi G, Schaefer AL, Mattmann ME, Christensen QH, Kessler A et al (2011) Isovaleryl-homoserine lactone, an unusual branched-chain quorum sensing signal from the soybean symbiont Bradyrhizobium japonicum. Proc Natl Acad Sci U S A 108:16765–16770
Christensen QH, Brecht RM, Dudekula D, Greenberg EP, Nagarajan R (2014) Evolution of acyl-substrate recognition by a family of acylhomoserine lactone synthases. PLoS One 9:e112464
Montebello AN, Brecht RM, Turner RD, Ghali M, Pu X, Nagarajan R (2014) Acyl-ACP substrate recognition in Burkholderia mallei BmaI1 acylhomoserine lactone synthase. Biochemistry 53:6231–6242
Gould TA, Schweizer HP, Churchill MEA (2002) Structure of the Pseudomonas aeruginosa acyl-homoserine lactone synthase LasI. Mol Microbiol 53:1135–1146
Gamage AM, Shui G, Wenk MR, Chua KL (2011) N-octanoylhomoserine lactone signaling mediated by BpsI-BpsR quorum sensing system plays a major role in biofilm formation of Burkholderia pseudomallei. Microbiology 157:1176–1186
Watson WT, Minogue TD, Val DL, Bodman SB, Churchill MEA (2002) Structural basis and specificity of acyl-homoserine lactone signal production in bacterial quorum sensing. Mol Cell 9:685–694
Duerkop BA, Herman JP, Ulrich RL, Churchill MEA, Greenberg EP (2008) The Burkholderia mallei BmaR3-BmaI3 quorum sensing system produces and responds to N-3-hydroxy-octanoyl homoserine lactone. J Bacteriol 190:5137–5141
Raychaudhari A, Jerga A, Tipton PA (2005) Chemical mechanism and substrate specificity of RhlI, an acylhomoserine lactone synthase from Pseudomonas aeruginosa. Biochemistry 44:2974–2981
Stadman ER (1957) Preparation and assay of acyl coenzymeA and other thiol esters: use of hydroxylamine. Methods Enzymol 3:931–946
Gutierrez JA, Crowder T, Rinaldo-Matthis A, Ho MC, Almo SC, Schramm VL (2009) Transition state analogues of 5′-methylthioadenosine nucleosidase disrupt quorum sensing. Nat Chem Biol 5:251–257
Quadri LE, Weinreb PH, Lei M, Nakano MM, Zuber P, Walsh CT (1998) Characterization of Sfp, a Bacillus subtilis phosphopantetheinyl transferase for peptide carrier protein domains in peptide synthetases. Biochemistry 37:1585–1595
La Clair JJ, Foley TL, Schegg TR, Regan CM, Burkart MD (2004) Manipulations of proteins in antibiotic synthesis. Chem Biol 11:195–201
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Shin, D., Nagarajan, R. (2018). Enzymatic Assays to Investigate Acyl-Homoserine Lactone Autoinducer Synthases. In: Leoni, L., Rampioni, G. (eds) Quorum Sensing. Methods in Molecular Biology, vol 1673. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7309-5_13
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7309-5_13
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7308-8
Online ISBN: 978-1-4939-7309-5
eBook Packages: Springer Protocols