A kinase innate process involves any drug induced change to the kinase itself which often makes it a better substrate for upstream activators or even a worse substrate for deactivating phosphatases. Countless protein kinase inhibitors have already been developed which don’t induce their target kinases to become hyperphosphorylated pan HDAC inhibitor to the activating sites. We carried out a double Akt transfection experiment as a further test with this type and to rule out any low catalytic activity mediated signals from Akt. The test utilizes the co transfection of banner wtAkt1 and HA asAkt1. Then only the Akt with the capacity of drug binding must be hyperphosphorylated, In the event the occupancy of the ATP website was the only determinant of hyperphosphorylation. In cells denver transfected with HA asAkt1 and flagwtAkt1, treatment with PrIDZ unveiled Thr308 and Ser473 phosphorylation is induced only on HA asAkt1 and not on drug insensitive banner wtAkt1 after immunoprecipitation. The finding shows that feedback Plastid mediated by signaling of Akt is not involved in hyperphosphorylation of Akt. The ability of flag tagged Akt1 to become hyperphosphorylated by Akt inhibitors was confirmed separately. An additional tagged build of asAkt1 containing mCherry, which indicates a big MW gel shift from endogenous Akt was also studied, with similar results. One prediction of the kinase intrinsic type of chemical caused Akt hyperphosphorylation is that drug binding should cause relocalization of Akt from the cytoplasm to the membrane. No known kinase inhibitors that we know about induce cellular translocation of their target kinase upon binding. To ascertain whether such a drug induced cellular relocalization was in reality occurring, we performed immunofluorescence studies of Akt. We chose to utilize untransfected HEK293 cells and A 443654, rather than asAkt LY2484595 transfected cells and PrIDZ, to prevent over-expression of the kinase. In particular, the cells maintain the stoichiometry between Akt and PIP3 although excess asAkt molecules may be mislocalized in asAkt overexpressed cells as a result of insufficient PIP3. Set cells were stained with anti pThr308 and anti Akt to determine the place of Akt and pAkt, after HEK293 cells were treated with A 443654. In the absence of any growth factor stimulation, treatment having A 443654 triggered translocation of Akt to the plasma membrane. More over, the membrane nearby Akt was phosphorylated at Thr308. In addition, both the occasions and the translocation were inhibited by pre-treatment with PIK90. Merck has noted an allosteric Akt inhibitor, Akti, which binds not in the active site and inhibits in vitro kinase activity. Curiously, in cells Akti also inhibits progress factor stimulated activation of Akt by preventing phosphorylation at Ser473 and Thr308 in a PH domain dependent manner.