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Abstract

The chemotaxis network, one of the most prominent prokaryotic sensory systems, is present in most motile bacteria and archaea. Although the conserved signaling core of the network is well characterized, ligand specificities of a large majority of diverse chemoreceptors encoded in bacterial genomes remain unknown. Here we performed a systematic identification and characterization of new chemoeffectors for the opportunistic pathogen Pseudomonas aeruginosa, which has 26 chemoreceptors possessing most of the common types of ligand binding domains. By performing capillary chemotaxis assays for a library of growth-promoting compounds, we first identified a number of novel chemoattractants of varying strength. We subsequently mapped specificities of these ligands by performing Forster resonance energy transfer (FRET) and microfluidic measurements for hybrids containing ligand binding domains of P. aeruginosa chemoreceptors and the signaling domain of the Escherichia coli Tar receptor. Direct binding of ligands to chemoreceptors was further confirmed in vitro using thermal shift assay and microcalorimetry. Altogether, the combination of methods enabled us to assign several new attractants, including methyl 4-aminobutyrate, 5-aminovalerate,L-ornithine, 2-phenylethylamine and tyramine, to previously characterized chemoreceptors and to annotate a novel purine-specific receptor PctP. Our screening strategy could be applied for the systematic characterization of unknown sensory domains in a wide range of bacterial species.Competing Interest StatementThe authors have declared no competing interest.