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

Autor: van der Wel T; Department of Molecular Physiology, Leiden Institute of Chemistry, Leiden University & Oncode Institute, Leiden, The Netherlands., Hilhorst R; PamGene International BV, 's-Hertogenbosch, The Netherlands., den Dulk H; Department of Molecular Physiology, Leiden Institute of Chemistry, Leiden University & Oncode Institute, Leiden, The Netherlands., van den Hooven T; PamGene International BV, 's-Hertogenbosch, The Netherlands., Prins NM; Department of Molecular Physiology, Leiden Institute of Chemistry, Leiden University & Oncode Institute, Leiden, The Netherlands., Wijnakker JAPM; Department of Molecular Physiology, Leiden Institute of Chemistry, Leiden University & Oncode Institute, Leiden, The Netherlands., Florea BI; Department of Bio-organic Synthesis, Leiden Institute of Chemistry, Leiden University, Leiden, The Netherlands., Lenselink EB; Department of Drug Discovery & Safety, Leiden Academic Centre for Drug Research, Leiden University, Leiden, The Netherlands., van Westen GJP; Department of Drug Discovery & Safety, Leiden Academic Centre for Drug Research, Leiden University, Leiden, The Netherlands., Ruijtenbeek R; PamGene International BV, 's-Hertogenbosch, The Netherlands., Overkleeft HS; Department of Bio-organic Synthesis, Leiden Institute of Chemistry, Leiden University, Leiden, The Netherlands., Kaptein A; Covalution Biosciences BV, Ravenstein, The Netherlands., Barf T; Covalution Biosciences BV, Ravenstein, The Netherlands., van der Stelt M; Department of Molecular Physiology, Leiden Institute of Chemistry, Leiden University & Oncode Institute, Leiden, The Netherlands. m.van.der.stelt@chem.leidenuniv.nl.
Jazyk: angličtina
Zdroj: Nature communications [Nat Commun] 2020 Jun 25; Vol. 11 (1), pp. 3216. Date of Electronic Publication: 2020 Jun 25.
DOI: 10.1038/s41467-020-17027-5
Abstrakt: 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.
Databáze: MEDLINE