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

A Screen for Membrane Fission Catalysts Identifies the ATPase EHD1.

Author information

1
Indian Institute of Science Education and Research , Dr. Homi Bhabha Road , Pashan, Pune 411008 , Maharashtra , India.

Abstract

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.

PMID:
30403133
PMCID:
PMC6327249
DOI:
10.1021/acs.biochem.8b00925
[Indexed for MEDLINE]
Free PMC Article

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