Figure 1

Activator-3, an AMPK mimetic is a potent pan-AMPK activator in cell based and cell free assays. (A) Chemical structures of Activator-3, AMP, ZMP and C2. Bottom panel: Rationale why Activator-3 may likely mimic AMP. (B) pAMPK based dose response curve of Activator-3 in primary rat hepatocytes (left panel) and rat L6 myoblasts (Right Panel). Results are expressed as the increase in activity relative to DMSO control and represent the mean ± SD for three independent experiments. (C) Human recombinant AMPK complexes (α1β1γ1, α2β1γ1, α2β2γ2 and α2β2γ3) expressed in baculo virus were assayed for allosteric activation by Activator-3. The representative four AMPK isozymes used for assays represent different combinations of all known reported isoforms like α1, α2, β1, β2, γ1, γ2 and γ3. EC₅₀ values were calculated by plotting a non-linear curve of log [agonist] vs. response. Results are expressed as the increase in activity relative to DMSO control and represent the mean ± SD for three independent experiments. (D) Enzymes allosterically regulated by AMP (PFK1) or using AMP as a substrate (AK) were assayed in presence of AMP (≤1 mM) or Activator-3 (≤10 µM). Both the enzymes were unaffected by Activator-3. Results are representative of three independent experiments conducted using commercially available kits.