Structure-kinetic relationship reveals the mechanism of selectivity of FAK inhibitors over PYK2

Cell Chem Biol. 2021 Jan 21;S2451-9456(21)00003-9. doi: 10.1016/j.chembiol.2021.01.003. Online ahead of print.


There is increasing evidence of a significant correlation between prolonged drug-target residence time and increased drug efficacy. Here, we report a structural rationale for kinetic selectivity between two closely related kinases: focal adhesion kinase (FAK) and proline-rich tyrosine kinase 2 (PYK2). We found that slowly dissociating FAK inhibitors induce helical structure at the DFG motif of FAK but not PYK2. Binding kinetic data, high-resolution structures and mutagenesis data support the role of hydrophobic interactions of inhibitors with the DFG-helical region, providing a structural rationale for slow dissociation rates from FAK and kinetic selectivity over PYK2. Our experimental data correlate well with computed relative residence times from molecular simulations, supporting a feasible strategy for rationally optimizing ligand residence times. We suggest that the interplay between the protein structural mobility and ligand-induced effects is a key regulator of the kinetic selectivity of inhibitors of FAK versus PYK2.

Keywords: NanoBRET; focal adhesion kinase (FAK); kinase inhibitor; ligand residence time; proline-rich tyrosine kinase 2 (PYK2); structure-kinetic-relationship; τRAMD.