A high-throughput screen for quorum-sensing inhibitors that target acyl-homoserine lactone synthases.
Many Proteobacteria use N-acyl-homoserine lactone (acyl-HSL) quorum sensing to control specific genes. Acyl-HSL synthesis requires unique enzymes that use S-adenosyl methionine as an acyl acceptor and amino acid donor. We developed and executed an enzyme-coupled high-throughput cell-free screen to discover acyl-HSL synthase inhibitors. The three strongest inhibitors were equally active against two different acyl-HSL synthases: Burkholderia mallei BmaI1 and Yersinia pestis YspI. Two of these inhibitors showed activity in whole cells. The most potent compound behaves as a noncompetitive inhibitor with a Ki of 0.7 µM and showed activity in a cell-based assay. Quorum-sensing signal synthesis inhibitors will be useful in attempts to understand acyl-HSL synthase catalysis and as a tool in studies of quorum-sensing control of gene expression. Because acyl-HSL quorum-sensing controls virulence of some bacterial pathogens, anti-quorum-sensing chemicals have been sought as potential therapeutic agents. Our screen and identification of acyl-HSL synthase inhibitors serve as a basis for efforts to target quorum-sensing signal synthesis as an antivirulence approach.
Department of Microbiology, University of Washington, Seattle, WA 98195, USA.
Quin H
QH
Christensen
Tyler L
TL
Grove
Squire J
SJ
Booker
E Peter
EP
Greenberg
D054742:Acyl-Butyrolactones / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000302:isolation & purification*; D004926:Escherichia coli; D005453:Fluorescence; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology*; D057166:High-Throughput Screening Assays; D020128:Inhibitory Concentration 50; D007700:Kinetics; D008025:Ligases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D015394:Molecular Structure; D010078:Oxazines; D020560:Proteobacteria / Q000502:physiology*; D053038:Quorum Sensing / Q000502:physiology*
D054742:Acyl-Butyrolactones; D004791:Enzyme Inhibitors; D010078:Oxazines; C014180:resorufin; D008025:Ligases
bacterial communication; cell–cell signaling; enzyme inhibitors