Tackling the bottleneck in bacterial signal transduction research: high-throughput identification of signal molecules.

Signal transduction processes are typically initiated by the interaction of signal molecules with sensor domains. The current lack of information on the signal molecules that feed into regulatory circuits forms a major bottleneck that hampers the understanding of regulatory processes. McKellar et al. report a high-throughput approach for the identification of signal molecules, which is based on thermal shift assays of recombinant sensor domains in the absence and presence of compounds from commercially available ligand collections. Initial binding studies with the sensor domain of the PctA chemoreceptor of Pseudomonas aeruginosa showed a close match between thermal shift assay results and microcalorimetric studies reported previously. Using thermal shift assays the authors then identify signals that bind to three chemoreceptors of the kiwifruit pathogen P. syringae pv. Actinidiae NZ-V13. Microcalorimetric binding studies and chemotaxis assays have validated the relevance of these ligands. The power of this technique lies in the combination of a high-throughput analytical tool with commercially available compound collections. The approach reported is universal since it can be employed to identify signal molecules to any type of sensor domain. There is no doubt that this technique will facilitate the identification of many signal molecules in future years.