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PreprintCited by 0Cellular transport and secretionNeuroscience and Neuropharmacology ResearchExtracellular vesicles in disease

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SNAP25 disease mutations change the energy landscape for synaptic exocytosis due to aberrant SNARE interactions

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Kádková, Anna, Murach, Jacqueline, Østergaard, Maiken, Malsam, Andrea, Malsam, Jörg, Lolicato, Fabio, Nickel, Walter, Söllner, Thomas H., Sørensen, Jakob B.·2024·eLife
DOI 10.7554/elife.88619.2

Preprint — published as 10.7554/elife.88619.3

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Abstract

Abstract SNAP25 is one of three neuronal SNAREs driving synaptic vesicle exocytosis. We studied three mutations in SNAP25 that cause epileptic encephalopathy: V48F, and D166Y in the Synaptotagmin-1 (Syt1) binding interface, and I67N, which destabilizes the SNARE-complex. All three mutations reduced Syt1-dependent vesicle docking to SNARE-carrying liposomes and Ca2+-stimulated membrane fusion in vitro and in neurons. The V48F and D166Y mutants (with potency D166Y > V48F) led to reduced Readily Releasable Pool (RRP) size, due to increased spontaneous (mEPSC) release and decreased priming rates. These mutations lowered the energy barrier for fusion and increased the release probability, which are gain-of-function features not found in Syt1 knockout (KO) neurons; normalized mEPSC release rates were higher (potency D166Y>V48F) than in the Syt1 KO. These mutations (potency D166Y > V48F) increased spontaneous association to partner SNAREs, resulting in unregulated membrane fusion. In contrast, the I67N mutant decreased mEPSC frequency and evoked EPSC amplitudes due to an increase in the apparent height of the energy barrier for fusion, whereas the RRP size was unaffected. This could be partly compensated by positive charges lowering the energy barrier. Overall, pathogenic mutations in SNAP25 cause complex changes in the energy landscape for priming and fusion.

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Metadata

Authors
Kádková, A., Murach, J., Østergaard, M., Malsam, A., Malsam, J., Lolicato, F., Nickel, W., Söllner, T. H., & Sørensen, J. B.
Affiliations
  • University of Copenhagen (DK)
  • Heidelberg University Biochemistry Center
Corresponding author
Sørensen, Jakob B.
ORCID
Venue
eLife
Publisher
eLife Sciences Publications, Ltd
Published
2024-01-19
ISSN
2050-084X (ISSN-L)
DOI
10.7554/elife.88619.2
OpenAlex
W4391034767
Cited by
0
References
6
Type
posted content
Language
en
Open access
Licence
Locations
eLife (acceptedVersion, cc-by)
Funders
  • Novo Nordisk Foundation (NNF19OC0058298)
  • Independent Research Fund Denmark (8020-00228A)
  • Lundbeck Foundation (R277-2018-802)
  • Deutsche Forschungsgemeinschaft (278001972 - TRR 186, LO 2821/1-1, SFB/TRR 186 A1, Ni 423/13-1, INST 35/1314-1 FUGG, INST 35/1503-1 FUGG)
  • CSC - IT Center for Science
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
Keywords
SNAP-25, SNARE complex, synaptic vesicle exocytosis, synaptotagmin-1, epileptic encephalopathy, readily releasable pool, spontaneous release, miniature excitatory postsynaptic currents, gain-of-function mutations

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