Two Pleckstrin homology domains scaffold SNAP-47-containing SNARE complexes
- 1University of Richmond
Abstract
Long-term potentiation requires the regulated insertion of AMPA receptors into the postsynaptic membrane of neurons, a process mediated by a SNARE complex containing SNAP-47, syntaxin-3, and synaptobrevin-2. Compared to other SNAP proteins, SNAP-47 exhibits slower fusion kinetics and contains uncharacterized regions whose functions remain unclear. Here, we integrated biochemical approaches with artificial intelligence structure prediction tools and molecular dynamics to investigate the architecture of SNAP-47-containing SNARE complexes. AlphaFold3 predictions of the ternary SNAP-47-containing SNARE complex revealed one known Pleckstrin homology domain (PH1) and one previously unidentified PH domain (PH2), which we validated experimentally. BioEmu and molecular dynamics simulations indicated that PH1 and PH2 form a stable and energetically favorable interface. In this bound state, PH1 stably interacted with the SNARE coiled coil, and in silico acetylation at a lysine whose acetylation is known to impair SNAP-47 function disrupted this PH1-SNARE interface. AMPAR insertion via SNAP-47-containing SNARE complexes is triggered by the calcium sensor synaptotagmin, and we observed that PH1, PH2, and synaptotagmin-1 can simultaneously engage with the plasma membrane, consistent with a pro-fusion conformation. Together, we provide new mechanistic insight into how PH domains regulate SNAP-47-mediated membrane fusion during synaptic plasticity.
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BibTeX
@misc{alickovic2026pleckstrin,
title = {{Two Pleckstrin homology domains scaffold SNAP-47-containing SNARE complexes}},
author = {I. Alickovic and M. Wise and R. Tela and C. Fausnaught and L. Lee and C. Beasley and C. Carr and C. Carr and L. Schwindt and J. J. Peters},
howpublished = {bioRxiv},
year = {2026},
month = oct,
doi = {10.64898/2026.10.15.759244},
url = {https://www.biorxiv.org/content/10.64898/2026.10.15.759244v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - Two Pleckstrin homology domains scaffold SNAP-47-containing SNARE complexes
AU - I. Alickovic
AU - M. Wise
AU - R. Tela
AU - C. Fausnaught
AU - L. Lee
AU - C. Beasley
AU - C. Carr
AU - C. Carr
AU - L. Schwindt
AU - J. J. Peters
PY - 2026
DA - 2026/10/15
PB - bioRxiv
DO - 10.64898/2026.10.15.759244
UR - https://www.biorxiv.org/content/10.64898/2026.10.15.759244v1
AB - Long-term potentiation requires the regulated insertion of AMPA receptors into the postsynaptic membrane of neurons, a process mediated by a SNARE complex containing SNAP-47, syntaxin-3, and synaptobrevin-2. Compared to other SNAP proteins, SNAP-47 exhibits slower fusion kinetics and contains uncharacterized regions whose functions remain unclear. Here, we integrated biochemical approaches with artificial intelligence structure prediction tools and molecular dynamics to investigate the architecture of SNAP-47-containing SNARE complexes. AlphaFold3 predictions of the ternary SNAP-47-containing SNARE complex revealed one known Pleckstrin homology domain (PH1) and one previously unidentified PH domain (PH2), which we validated experimentally. BioEmu and molecular dynamics simulations indicated that PH1 and PH2 form a stable and energetically favorable interface. In this bound state, PH1 stably interacted with the SNARE coiled coil, and in silico acetylation at a lysine whose acetylation is known to impair SNAP-47 function disrupted this PH1-SNARE interface. AMPAR insertion via SNAP-47-containing SNARE complexes is triggered by the calcium sensor synaptotagmin, and we observed that PH1, PH2, and synaptotagmin-1 can simultaneously engage with the plasma membrane, consistent with a pro-fusion conformation. Together, we provide new mechanistic insight into how PH domains regulate SNAP-47-mediated membrane fusion during synaptic plasticity.
ER -