Chemical genetics strategy to profile kinase target engagement reveals role of FES in neutrophil phagocytosis.
Tom van der WelRiet HilhorstHans den DulkTim van den HoovenNienke M PrinsJoost A P M WijnakkerBogdan I FloreaEelke B LenselinkGerard van WestenRob RuijtenbeekHerman S OverkleeftAllard KapteinTjeerd BarfMario van der SteltPublished in: Nature communications (2020)
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.
Keyphrases
- tyrosine kinase
- induced apoptosis
- crispr cas
- drug discovery
- epidermal growth factor receptor
- social media
- cell cycle arrest
- living cells
- genome editing
- protein kinase
- cell death
- emergency department
- liver failure
- cell therapy
- bone marrow
- drug induced
- machine learning
- big data
- hepatitis b virus
- deep learning
- small molecule
- genome wide identification
- genome wide analysis