Enzymatic assays to investigate acyl-homoserine lactone autoinducer synthases

Daniel Shin, Rajesh Nagarajan

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

1 Scopus citations

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.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages161-176
Number of pages16
DOIs
StatePublished - 2018

Publication series

NameMethods in Molecular Biology
Volume1673
ISSN (Print)1064-3745

Keywords

  • 5′-Deoxy-5′-(methylthio)adenosine
  • Acyl carrier protein
  • AHL synthase
  • Coenzyme A
  • Colorimetric assay
  • HPLC assay
  • Methylthioadenosine nucleosidase
  • N-acyl-homoserine lactone
  • Quorum sensing
  • S-adenosyl-L-methionine
  • Spectrophotometric assay
  • Xanthine oxidase

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