Hunter, R., Foretz, M., Bultot, L., Fullerton, M., Deak, M., Ross, F., Hawley, S., Shpiro, N., Viollet, B., Barron, D., Kemp, B., Steinberg, G., Hardie, D. G & Sakamoto, K. (2014). Mechanism of action of compound-13 : An a1-selective small molecule activator of AMPK. Chemistry and Biology,21(7), 866-879. Retrieved from https://doi.org/10.1016/j.chembiol.2014.05.014
AMPK is a sensor of cellular energy status and a promising target for drugs aimed at metabolic disorders. We have studied the selectivity and mechanism of a recently described activator, C2, and its cell-permeable prodrug, C13. C2 was a potent allosteric activator of α1-complexes that, like AMP, also protected against Thr172 dephosphorylation. Compared with AMP, C2 caused only partial allosteric activation of α2-complexes and failed to protect them against dephosphorylation. We show that both effects could be fully restored by exchanging part of the linker between the autoinhibitory and C-terminal domains in α2, containing the equivalent region from α1 thought to interact with AMP bound in site 3 of the γ subunit. Consistent with our results in cell-free assays, C13 potently inhibited lipid synthesis in hepatocytes from wild-type and was largely ineffective in AMPK-knockout hepatocytes; its effects were more severely affected by knockout of α1 than of α2, β1, or β2.