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. 2019 Jan 8;39(1):BSR20181252. doi: 10.1042/BSR20181252

Figure 1. Effects of Rabs on Myo5b ATPase activity.

Figure 1

(A) Schematic primary structure of Myo5b and its binding sites with Rabs. Myo5b-FL, the full-length Myo5b; IQ, the CaM binding site; GTD, the C-terminal globular tail domain. (B) Anti-Flag pull down assays reveal the interaction of Myo5b tail with GST–Rab8a, 10 and 11a. (C) Anti-Flag pull down of Flag-Myo5b tail constructs with GST-Rabs. (D) Effects of Rabs on Myo5b ATPase activity. The ATPase activity was measured in the presence of ∼0.1 μM Myo5b-FL, 40 μM actin and 12 μM GST-Rab in a solution containing 20 mM MOPS-KOH (pH 7.0), 200 mM KCl, 1 mM MgCl2, 1 mM DTT, 0.25 mg/ml BSA, 12 μM CaM, 0.5 mM ATP, 2.5 mM PEP, 20 U/ml pyruvate kinase and 1 mM EGTA. Control shows the ATPase activity of Myo5b-FL in the absence of GST-Rab. Values are mean ± S.D. from at least four independent assays; ***P<0.001. Anti-Flag pull down assays were repeated three times and typical results are presented.