Figure 5.

In vitro methylation assays characterizing AgCARM1 automethylation and its affect on enzyme activity. ³H-AdoMet was incubated with purified protein followed by SDS-PAGE, coomassie staining, and exposure to film. (A) In vitro methylation of known CARM1 substrates by AgCARM1^WT and AgCARM1^R485K. (B) Isolated and mixed automethylation reactions with 0.1 μg and/or 1 μg of AgCARM1. Top shows a cartoon representation of reaction contents and possible interactions. Looping arrows indicate possible intramolecular and/or intermolecular automethylation. Straight arrows indicate possible intermolecular automethylation. Bottom shows in vitro methylation results of these reaction conditions, with lane numbers corresponding to the same numbered cartoon above. *Band copurified with PABP1, likely due to degradation. CS, Coomassie Stain; AR, Autoradiograph.