Chemical genetics strategy to profile kinase target engagement reveals role of FES in neutrophil phagocytosis.

Nature communications 2020 Jun 25

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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

Tom van der Wel, Riet Hilhorst, Hans den Dulk, Tim van den Hooven, Nienke M Prins, Joost A P M Wijnakker, Bogdan I Florea, Eelke B Lenselink, Gerard J P van Westen, Rob Ruijtenbeek, Herman S Overkleeft, Allard Kaptein, Tjeerd Barf, Mario van der Stelt. Chemical genetics strategy to profile kinase target engagement reveals role of FES in neutrophil phagocytosis. Nature communications. 2020 Jun 25;11(1):3216