A high-throughput platform for real-time analysis of membrane fission reactions reveals dynamin function

Nat Cell Biol. 2015 Dec;17(12):1588-96. doi: 10.1038/ncb3254. Epub 2015 Oct 19.


Dynamin, the paradigmatic membrane fission catalyst, assembles as helical scaffolds that hydrolyse GTP to sever the tubular necks of clathrin-coated pits. Using a facile assay system of supported membrane tubes (SMrT) engineered to mimic the dimensions of necks of clathrin-coated pits, we monitor the dynamics of a dynamin-catalysed tube-severing reaction in real time using fluorescence microscopy. We find that GTP hydrolysis by an intact helical scaffold causes progressive constriction of the underlying membrane tube. On reaching a critical dimension of 7.3 nm in radius, the tube undergoes scission and concomitant splitting of the scaffold. In a constant GTP turnover scenario, scaffold assembly and GTP hydrolysis-induced tube constriction are kinetically inseparable events leading to tube-severing reactions occurring at timescales similar to the characteristic fission times seen in vivo. We anticipate SMrT templates to allow dynamic fluorescence-based detection of conformational changes occurring in self-assembling proteins that remodel membranes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Catalysis
  • Cell Membrane / chemistry
  • Cell Membrane / metabolism*
  • Cell Membrane / ultrastructure
  • Coated Pits, Cell-Membrane / chemistry
  • Coated Pits, Cell-Membrane / metabolism
  • Dynamin I / chemistry
  • Dynamin I / genetics
  • Dynamin I / metabolism*
  • Fluorescence Recovery After Photobleaching
  • Guanosine Triphosphate / chemistry
  • Guanosine Triphosphate / metabolism*
  • Humans
  • Hydrolysis
  • Membrane Lipids / chemistry
  • Membrane Lipids / metabolism
  • Microscopy, Confocal
  • Microscopy, Electron, Scanning / methods
  • Microscopy, Fluorescence
  • Models, Chemical
  • Models, Molecular
  • Molecular Conformation
  • Mutation
  • Phosphatidylinositol 4,5-Diphosphate / chemistry
  • Phosphatidylinositol 4,5-Diphosphate / metabolism
  • Protein Conformation
  • Time-Lapse Imaging / methods*


  • Membrane Lipids
  • Phosphatidylinositol 4,5-Diphosphate
  • Guanosine Triphosphate
  • Dynamin I