Chemical tools to monitor drug-target engagement of endogenously expressed protein kinases are highly desirable for preclinical target validation in drug discovery. Here, we describe a chemical genetics strategy to selectively study target engagement of endogenous kinases. By substituting a serine residue into cysteine at the DFG-1 position in the ATP-binding pocket, we sensitize the non-receptor tyrosine kinase FES towards covalent labeling by a complementary fluorescent chemical probe. This mutation is introduced in the endogenous FES gene of HL-60 cells using CRISPR/Cas9 gene editing. Leveraging the temporal and acute control offered by our strategy, we show that FES activity is dispensable for differentiation of HL-60 cells towards macrophages. Instead, FES plays a key role in neutrophil phagocytosis via SYK kinase activation. This chemical genetics strategy holds promise as a target validation method for kinases.
CITATION STYLE
van der Wel, T., Hilhorst, R., den Dulk, H., van den Hooven, T., Prins, N. M., Wijnakker, J. A. P. M., … van der Stelt, M. (2020). Chemical genetics strategy to profile kinase target engagement reveals role of FES in neutrophil phagocytosis. Nature Communications, 11(1). https://doi.org/10.1038/s41467-020-17027-5
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