FIG. 4.

The SSR of HK1S interacts with PFKM in a GST pull-down and coimmunoprecipitation assays. A) Diagram of recombinant proteins containing either TSR of PFKMS fused to GST (GST-TSR) or all or part of PFKM fused to GST, including full length (GST-PFKM-FL), the N-terminal region, and the C-terminal region (GST-PFKM-N and GST-PFKM-C). B) GST-PFKM-FL, GST-PFKM-N, and GST-PFKM-C proteins were immobilized on glutathione-sepharose resin and incubated with FLAG-tagged SSR protein, and the recombinant proteins that interacted were eluted from the resin. The eluted recombinant proteins were separated by SDS-PAGE, and then Western blot analysis was performed with antibodies to FLAG (upper panel) or GST (lower panel). C) In vitro-translated [³⁵S]-HK1S added to testis lysate coimmunoprecipitated with PFKMS when TSR antiserum and protein G beads were added but not when TSR antiserum was deleted (-) or rabbit IgG was added instead of TSR antiserum.