FIGURE 7.

Endogenously tagged TbPRMT7 exhibits type III PRMT activity. A, endogenously tagged TbPRMT7 has in vitro activity comparable to recombinant GST-TbPRMT7. Right panel, the indicated substrates were incubated with 100 ng of purified TbPRMT7-ProtC from T. brucei incubated with either PBS buffer alone (-), 3 μg of RBP16, or 10 μg of bovine histone in the presence of 2 μCi of [³H]AdoMet in PBS overnight at room temperature. Left panel, three micrograms of GST-PRMT7 was used as a positive control with each substrate for activity in vitro. For each reaction the Coomassie Blue stain of the substrate is shown below the resultant fluorograph of activity. B, purified TbPRMT7-ProtC from T. brucei still exhibits Type III PRMT activity. The reactions from A were subjected to high resolution cation exchange chromatography as in Fig. 3.