Fig. 3. AD80 is active against gatekeeper mutant RETV804M cells.

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Fig. 3. AD80 is active against gatekeeper mutant RETV804M cells. AD80 is active against gatekeeper mutant RETV804M cells. (A) Optimized structures after extensive MD refinement followed by ALPB optimization. (i) RETwt/AD80 after 102 ns, (ii) RETwt/AD57 after 202 ns (92 ns from RETwt/AD80 simulation followed by 110 ns from TI-MD), and (iii) RETV804M/AD80 after 107 ns (side view). The DFG motif is shown in violet. Distances from the center of central phenyl to Val804-C(wt), Ile788-C(wt), and Met804-S(V804M) are 4.77, 3.90, and 4.29 Å, respectively. Dashed lines indicate the H-bond between the bound ligands and aspartate of the DFG motif. (B) Heat map of mean 50% growth inhibition (GI50) values (n ≥ 3) of Ba/F3 cells expressing CCDC6-RETV804M or KIF5B-RETV804M after 72 hours of treatment, as assessed for various inhibitors. (C) Immunoblotting of AD80-, cabozantinib-, or vandetanib-treated (4 hours) KIF5B-RETV804M Ba/F3 cells. (D) Immunoblotting of Ba/F3 cells expressing CCDC6-RET-RETwt or CCDC6-RETV804M under AD80 or vandetanib treatment (4 hours). wt, wild type. (E) Calculated Michaelis constant (Km) values of ATP binding to RETwt or RETV804M from three independent experiments. ***P < 0.001, n = 3. Dennis Plenker et al., Sci Transl Med 2017;9:eaah6144 Copyright © 2017 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.