FIG. 2.

CK2 binding to EB2. (A) Schematic representation of EB2 and EB2 polypeptides used to generate GST fusion proteins. NLS, nuclear localization signal; RBD, RNA-binding domain. (B) Recombinant CK2α and CK2β in vitro labeled with [³⁵S]methionine (lanes 1 to 3), either individually (lanes 4 to 11) or mixed together (lanes 12 to 15), for 15 min at 4°C were incubated with glutathione-Sepharose 4B beads loaded with GST or GST-A, -Nter, or -Cter. Proteins bound to the beads were eluted, resolved by SDS-PAGE, and visualized by Coomassie blue staining (bottom, lanes 4 to 15) and by autoradiography (top, lanes 4 to 15). *, nonspecific band. (C) Purified CK2, ³²P labeled by autophosphorylation in vitro (b, lane 1), was incubated with beads loaded with GST or GST-A, -B, -C, and -Nter. Proteins bound to the beads were eluted, resolved by SDS-PAGE, and visualized by Coomassie blue staining (a) and by autoradiography (b).