1. Interaction with PRMT5 involves the RBD1,2 and RGG domains of nucleolin.
Interaction with PRMT5 involves the RBD1,2 and RGG domains of nucleolin. A, schematic representation of nucleolin and the different recombinant proteins produced in E. coli for use in these interaction studies. Polypeptides representing the RNA binding domains (RBD1,2) and/or the arginine/glycine-rich domain (RGG) of nucleolin fused to MBP tag at their NH2 termini were used. B, whole-cell lysate from DU145 cells was incubated with purified MBP-tagged polypeptides, which were then captured using amylose-linked magnetic beads. After extensive washing, bound proteins were eluted and analyzed by Western blotting for with anti-PRMT5 (top) and anti-MBP, as a control for loading (bottom). As a negative control, purified MBP-tagged RGG was incubated with amylose beads in the absence of cell lysate (lane 5).
Yun Teng et al. Cancer Res 2007;67:
©2007 by American Association for Cancer Research