Figure 5
Interaction network for proteins associated with AMPK complex in regulation of actin cytoskeleton.
(A) Network analysis of AMPK interactome, showing that the AMPK complex interacts directly with multiple proteins associated with the regulation of the actin cytoskeleton. Proteins and protein-protein interactions associated with regulation of the actin cytoskeleton were extracted from the KEGG pathway and STRING 9.1 databases. All input proteins associated with actin regulation are depicted as small gray spheres. Interacting proteins in our dataset are depicted as green spheres and interacting proteins validated by Co-IP are shown as yellow spheres. Bait proteins are shown as red hexagons. Protein-protein interactions extracted from the STRING database are shown as grey lines. Interactions identified in this study are shown as green lines. Co-IP-validated interactions are shown as red dotted lines. (B) Validation of protein-protein interactions by pulldown approach. A novel interacting protein (Iqgap1) was validated by co-IP. Well-known proteins (Gsn and Vim) were also validated. Vim was used as a positive control to confirm homologous interactions. (C) Validation of protein-protein interactions using direct IP. AMPK-α1, AMPK-β1, Myh9, Iqgap1, Rhoa and Rac1 were validated by co-IP. The co-IP results for novel interactors (Myh9, Iqgap1, Rhoa and Rac1) that were associated with regulation of the actin cytoskeleton corresponded well with the AP-MS results.
