Protein associated with myc (PAM) is a giant E3 ubiquitin ligase of 510 kDa. While the role of PAM during neuronal development is well established, very little is known about its function in the regulation of synaptic strength. Here we employed Multi-Epitope-Ligand-Carthography (MELC) to study protein network profiles associated with PAM during the modulation of synaptic strength. MELC is a novel imaging technology that utilizes biomathematical tools to describe protein networks after consecutive immunohistochemical visualization of up to 100 proteins on the same sample. As in vivo model to modulate synaptic strength we used the formalin test, a common model for acute and inflammatory pain. MELC analysis was performed with 37 different antibodies or fluorescence tags on spinal cord slices and led to the identification of 1390 PAM-related motifs that distinguish untreated and formalin-treated spinal cords. The majority of these motifs related to ubiquitin-dependent processes and/or the actin cytoskeleton. We detected an intermittent colocalisation of PAM and ubiquitin with TSC2, a known substrate of PAM, and the glutamate receptors mGluR5 and GluR1. Importantly these complexes were detected exclusively in the presence of F-actin. A direct PAM/F-actin interaction was confirmed by colocalisation and cosedimentation. The binding of PAM towards F-actin varied strongly between the PAM splice forms found in rat spinal cords. PAM did not ubiquitylated actin or altered actin polymerization and depolymerization. However, F-actin decreased the ubiquitin ligase activity of purified PAM. Since PAM-activation is known to involve its translocation, the binding of PAM to F-actin may serve to control its subcellular localization as well as its activity. Taken together we show that defining protein network profiles by topological proteomic analysis is a useful tool to identify previously unknown protein-protein interactions that underlie synaptic processes.