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

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 and Mario van der Stelt ()
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Tom van der Wel: Leiden Institute of Chemistry, Leiden University & Oncode Institute
Riet Hilhorst: PamGene International BV
Hans den Dulk: Leiden Institute of Chemistry, Leiden University & Oncode Institute
Tim van den Hooven: PamGene International BV
Nienke M. Prins: Leiden Institute of Chemistry, Leiden University & Oncode Institute
Joost A. P. M. Wijnakker: Leiden Institute of Chemistry, Leiden University & Oncode Institute
Bogdan I. Florea: Leiden Institute of Chemistry, Leiden University
Eelke B. Lenselink: Leiden Academic Centre for Drug Research, Leiden University
Gerard J. P. van Westen: Leiden Academic Centre for Drug Research, Leiden University
Rob Ruijtenbeek: PamGene International BV
Herman S. Overkleeft: Leiden Institute of Chemistry, Leiden University
Allard Kaptein: Covalution Biosciences BV
Tjeerd Barf: Covalution Biosciences BV
Mario van der Stelt: Leiden Institute of Chemistry, Leiden University & Oncode Institute

Nature Communications, 2020, vol. 11, issue 1, 1-20

Abstract: Abstract 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.

Date: 2020
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DOI: 10.1038/s41467-020-17027-5

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