Figure 2.

Hydrophobic VL1-membrane interactions are critical for dynamin function. (A) Kinetics of transferrin uptake in cells expressing Dyn1 WT (●), Dyn1 I533W (▴), Dyn1 I533L (▵), or Dyn1 I533A (◊) were determined and plotted as in Figure 1A. (B) Lipid-stimulated GTPase activities of 0.5 μM Dyn1 WT (●), Dyn1 I533W (▴), Dyn1 I533L (▵), or Dyn1 I533A (◊) preincubated with PIP₂-containing liposomes (150 μM total lipid) were obtained and plotted as in Figure 1B. (C) Sedimentation analysis of 1.0 μM Dyn1 I533A on PIP₂-containing liposomes (150 μM total lipid) was performed and plotted as in Figure 1C. (D) Membrane fission activities of 0.5 μM Dyn1 WT and Dyn1 I533A was determined on SUPER templates (6 μM total lipid) in the constant presence of GTP (1 mM) by a low-speed sedimentation assay as in Figure 1D.