A Screen for Membrane Fission Catalysts Identifies the ATPase EHD1

Biochemistry. 2019 Jan 8;58(1):65-71. doi: 10.1021/acs.biochem.8b00925. Epub 2018 Nov 16.


Membrane fission manifests during cell division, synaptic transmission, vesicular transport, and organelle biogenesis, yet identifying proteins that catalyze fission remains a challenge. Using a facile and robust assay system of supported membrane tubes in a microscopic screen that directly monitors membrane tube scission, we detect robust GTP- and ATP-dependent as well as nucleotide-independent fission activity in the brain cytosol. Using previously established interacting partner proteins as bait for pulldowns, we attribute the GTP-dependent fission activity to dynamin. Biochemical fractionation followed by mass spectrometric analyses identifies the Eps15-homology domain-containing protein1 (EHD1) as a novel ATP-dependent membrane fission catalyst. Together, our approach establishes an experimental workflow for the discovery of novel membrane fission catalysts.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biological Transport
  • Brain / metabolism*
  • Cell Membrane / chemistry*
  • Cell Membrane / metabolism*
  • Cytosol / metabolism*
  • Dynamins / metabolism*
  • Goats
  • Guanosine Triphosphate / metabolism*
  • High-Throughput Screening Assays
  • Humans
  • Vesicular Transport Proteins / metabolism*


  • EHD1 protein, human
  • Vesicular Transport Proteins
  • Guanosine Triphosphate
  • Dynamins