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Figure 3 | BMC Neuroscience

Figure 3

From: Kalirin12 interacts with dynamin

Figure 3

The IgFn domain of Kalirin interacts with the GTPase domain of dynamin. A. The GFP-tagged dynamin2 proteins expressed transiently in pEAK Rapid cells are shown diagrammatically; dynamin domains were defined in Fig. 1C. B.–E. Lysates were incubated with beads to which 2 μg GST (G) or 2 μg GST-IgFn (Ig) was bound. Bound proteins (50% of total) were eluted and fractionated on polyacrylamide gels; inputs (In; 1% of total) were analyzed simultaneously. EGFP and dynamin were visualized using antibody to GFP. B. Dynamin2 (Dyn2), but not dynamin2 K44A bound to GST-IgFn. The Coomassie Brilliant Blue image demonstrates similar amounts of GST and GST-IgFn. C. The GTPase domain of dynamin2 bound to Kalirin-IgFn. D. Binding of dynamin2 to GST-IgFn was enhanced by 1 mM GTP or 1 mM GTPγs. E. Dynamin2 lacking the PRD (ΔPRD) bound to Kalirin-IgFn while the GED/PRD domains of dynamin2 did not. F. pEAK Rapid cells expressing the dynamin2/GTPase-GFP fusion protein along with Kalirin7 or Kalirin12 were extracted and immunoprecipitated using a polyclonal GFP antibody or control immunoglobulin; the Kalirin-spectrin antibody detected an interaction of dynamin2 with Kalirin12, but not with Kalirin7.

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