Figure 7. The allosteric drug-induced Akt inhibited form is distinct from its native autoinhibited form.
(A) Binding assays of phospholipid PIP3 with full-length Akt in presence of 20 µM MK2206. Chemical structure of MK2206 is shown on the right. The fluorescence anisotropy measurements (n = 2) were carried out and fit to quadratic binding isotherms, and Kd values shown ± S.D. (B) Cartoon model illustrated the distinct conformational structure of PH domain of allosteric drug-bound Akt when compared to that of natural autoinhibited form.
Figure 7—figure supplement 1. Allosteric inhibitors glue PH and kinase domains together and compete with PIP3.
(A) Binding assays of soluble phospholipid PIP3 with full-length Akt in presence of 10 µM compound VIII (compd. VIII), fluorescence anisotropy measurements (n = 2) were carried out and fit to quadratic binding isotherms, and Kd values shown ± S.D. (B–C) MST measurements (n = 2) for trans-binding of varied amounts of PH domain with N-terminally Cy5-labeled Akt (aa 122–480) fragment without (B) and with 20 µM MK2206 (C), the obtained Kd values expressed in the inset boxes, S.D. shown.