Details

Autor(en) / Beteiligte

• van der Wel, Tom
• Hilhorst, Riet
• den Dulk, Hans
• van den Hooven, Tim
• Prins, Nienke M.
• Wijnakker, Joost A. P. M.
• Florea, Bogdan I.
• Lenselink, Eelke B.
• van Westen, Gerard J. P.
• Ruijtenbeek, Rob
• Overkleeft, Herman S.
• Kaptein, Allard
• Barf, Tjeerd
• van der Stelt, Mario

Titel

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

Ist Teil von

• Nature communications, 2020-06, Vol.11 (1), p.3216-3216, Article 3216

Ort / Verlag

London: Nature Publishing Group

Erscheinungsjahr

2020

Quelle

Free E-Journal (出版社公開部分のみ）

Beschreibungen/Notizen

• 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.