@article{a7ef884764bf4b8ca4fa5ba4b0681c04,
title = "Structure-kinetic relationship reveals the mechanism of selectivity of FAK inhibitors over PYK2",
abstract = "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 = "focal adhesion kinase (FAK), kinase inhibitor, ligand residence time, NanoBRET, proline-rich tyrosine kinase 2 (PYK2), structure-kinetic-relationship, τRAMD",
author = "Berger, \{Benedict Tilman\} and Marta Amaral and Kokh, \{Daria B.\} and Ariane Nunes-Alves and Djordje Musil and Timo Heinrich and Martin Schr{\"o}der and Rebecca Neil and Jing Wang and Iva Navratilova and Joerg Bomke and Elkins, \{Jonathan M.\} and Susanne M{\"u}ller and Matthias Frech and Wade, \{Rebecca C.\} and Stefan Knapp",
note = "Funding Information: This work was supported by a Capes-Humboldt postdoctoral scholarship to A.N.-A. (Capes process number 88881.162167/2017-01), the European Union's Horizon 2020 Framework Program for Research and Innovation under the Specific Grant Agreement no. 785907 and no. 945539 (Human Brain Project SGA2 and SGA3 to R.C.W.), EU/EFPIA Innovative Medicines Initiative Joint Undertaking, K4DD (grant no. 115366 to R.C.W. and S.K.) and the Klaus Tschira Foundation (D.B.K. A.N.-A. R.C.W.). B.-T.B. M.S. J.M.E. D.B.K. and S.K. are supported by the Innovative Medicines Initiative 2 Joint Undertaking under grant agreement no. 875510 and the SGC, a registered charity (number 1097737) that receives funds from AbbVie, Bayer AG, Boehringer Ingelheim, Canada Foundation for Innovation, Eshelman Institute for Innovation, Genentech, Genome Canada through Ontario Genomics Institute [OGI-196], EU/EFPIA/OICR/McGill/KTH/Diamond, Innovative Medicines Initiative 2 Joint Undertaking (EUbOPEN grant 875510), Janssen, Merck KGaA (a.k.a. EMD in Canada and USA), Merck (a.k.a. MSD outside Canada and USA), Pfizer, S?o Paulo Research Foundation-FAPESP, Takeda, and Wellcome. B.-T.B. and S.K. would like to acknowledge funding by the DFG-funded CRC SFB 1399. S.K. S.M. M.F. J.B. J.M.E. and R.C.W. supervised the research. T.H. synthesized the inhibitors. B.-T.B, M.S. D.M. and M.A. performed structural studies. B.-T. B. M.A. I.N. and J.W. performed biochemical and biophysical binding studies. B.-T.B. and M.A. performed NanoBRET cell-based ligand studies. D.B.K. A.N.-A. R.N. and R.C.W. performed in silico analysis. S.K. M.A. D.B.K. and B-T.B. drafted the manuscript, which has been edited and approved by all authors. M.F. J.B. and T.H. are employees of Merck KGaA, Darmstadt, Germany. M.A. was an employee of Merck KGaA, Darmstadt, Germany at the time of experimental procedures and is now an employee of Sanofi-Aventis Deutschland GmbH, Frankfurt, Germany. Other authors have no conflict of interest to declare. Funding Information: This work was supported by a Capes-Humboldt postdoctoral scholarship to A.N.-A. (Capes process number 88881.162167/2017-01 ), the European Union's Horizon 2020 Framework Program for Research and Innovation under the Specific Grant Agreement no. 785907 and no. 945539 (Human Brain Project SGA2 and SGA3 to R.C.W.), EU/EFPIA Innovative Medicines Initiative Joint Undertaking, K4DD (grant no. 115366 to R.C.W. and S.K.) and the Klaus Tschira Foundation (D.B.K., A.N.-A., R.C.W.). B.-T.B., M.S., J.M.E., D.B.K., and S.K. are supported by the Innovative Medicines Initiative 2 Joint Undertaking under grant agreement no. 875510 and the SGC , a registered charity (number 1097737 ) that receives funds from AbbVie , Bayer AG , Boehringer Ingelheim , Canada Foundation for Innovation , Eshelman Institute for Innovation , Genentech , Genome Canada through Ontario Genomics Institute [OGI-196], EU/EFPIA/OICR/McGill/KTH/Diamond , Innovative Medicines Initiative 2 Joint Undertaking ( EUbOPEN grant 875510), Janssen , Merck KGaA (a.k.a. EMD in Canada and USA), Merck (a.k.a. MSD outside Canada and USA), Pfizer , S{\~a}o Paulo Research Foundation- FAPESP , Takeda , and Wellcome . B.-T.B. and S.K. would like to acknowledge funding by the DFG -funded CRC SFB 1399. Publisher Copyright: {\textcopyright} 2021 Elsevier Ltd Copyright: Copyright 2021 Elsevier B.V., All rights reserved.",
year = "2021",
month = may,
day = "20",
doi = "10.1016/j.chembiol.2021.01.003",
language = "English",
volume = "28",
pages = "686--698.e7",
journal = "Cell Chemical Biology",
issn = "2451-9456",
publisher = "Elsevier",
number = "5",
}