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. 2009 Nov 15;20(22):4630–4639. doi: 10.1091/mbc.E09-08-0683

Figure 3.

Figure 3.

I533A VL1 mutant is defective in membrane curvature generation and membrane fission. (A) Fluorescence images of RhPE-labeled SUPER templates showing the effect of addition of either 0.5 μM Dyn1 WT or Dyn1 I533A in the absence of GTP. Images were acquired at a focal plane close to the coverslip. Contrast inverted images are also shown (right panels) for clarity. Note the absence of optically visible membrane tubules for Dyn1 I533A. (B) Fluorescence images showing the effect of addition of either 0.5 μM Dyn1 WT or Dyn1 I533A in the constant presence of GTP to membrane tubules (typically 0.5–1.0 μm in diameter) transiently generated by glycerol flow from RhPE-labeled SUPER templates (Pucadyil and Schmid, 2008). Left and middle, note the time-dependent release of vesicles (arrows) from the templates only in the case of Dyn1 WT. See Supplemental Movie 1 for entire sequence. Right, magnified fluorescence images showing visible membrane constrictions (arrows) before fission of flow-generated membrane tubules only in the case of Dyn1 WT.