A Screen for Membrane Fission Catalysts Identifies
the ATPase EHD1
Version 2 2018-11-17, 00:43
Version 1 2018-11-15, 21:14
Posted on 2018-11-17 - 00:43
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.
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Kamerkar, Sukrut
C.; Roy, Krishnendu; Bhattacharyya, Soumya; Pucadyil, Thomas J. (2018). A Screen for Membrane Fission Catalysts Identifies
the ATPase EHD1. ACS Publications. Collection. https://doi.org/10.1021/acs.biochem.8b00925
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AUTHORS (4)
SK
Sukrut
C. Kamerkar
KR
Krishnendu Roy
SB
Soumya Bhattacharyya
TP
Thomas J. Pucadyil
KEYWORDS
catalyze fissionnucleotide-independent fission activitymonitors membrane tube scissionBiochemical fractionationEps 15-homology domain-containing protein 1GTP-dependent fission activityassay systemnovel ATP-dependent membrane fission catalystbrain cytosolorganelle biogenesisATPase EHD 1 Membrane fission manifestsMembrane Fission Catalysts Identifiespartner proteinsnovel membrane fission catalystscell divisionsynaptic transmissionmembrane tubes