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. 2011 Feb 23;286(16):14352–14361. doi: 10.1074/jbc.M110.187286

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© 2011 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 2. — Rab3A is a novel Myo5a binding partner. A, synaptosomal fraction (Syn) and bound proteins (BP) eluted from immobilized GST-Myo5a tail affinity column were analyzed by SDS-PAGE stained with Coomassie. GST-Myo5a tail (84 kDa) and GST (25 kDa) are indicated. B, Western blots of the synaptosomal fraction (Syn) and bound proteins (BP) eluted from GST-Myo5a tail affinity column are shown. Blots were probed for proteins as indicated. Rab3A was detected in the eluted fraction. C-D, Rab3A and Rabphilin3A bind to Myo5a tail in a GTP-dependent manner. Inactive GDP-bound Rab3A (C, left panel) and Rabphilin3A (D, left panel) bind to RabGDI. Active GTP-bound Rab3A (C, right panel) and Rabphilin3A (D, right panel) bind to Myo5a tail in the presence of GTPγS. Signal intensities of Rab3A and Rabphilin3A were individually normalized against the signal obtained with these proteins in synaptosomes. All data are mean ± S.D. of three independent experiments. Statistically significant differences at p < 0.001 are indicated by asterisks (*).