Chemical genetics strategy to profile kinase target engagement reveals role of FES in neutrophil phagocytosis.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 32587248.
- Also identified by DOI 10.1038/s41467-020-17027-5 and PMC identifier 7316778.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
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.
Medical subject headings
- Fluorescence Resonance Energy Transfer
- Fluorescent Dyes
- Proto-Oncogene Proteins c-fes