The Interaction of Munc18-1 Helix 11 and 12 with the Central Region of the VAMP2 SNARE Motif Is Essential for SNARE Templating and Synaptic Transmission
| Autor: | Marieke Meijer, Susanne Kreye, Robert B. Russell, Matthew J. Betts, Philipp Brenner, Thomas H. Söllner, Timon André, Matthijs Verhage, Bernhard Dörr, Jessica Classen, Birte Zuidinga |
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| Přispěvatelé: | Human genetics, Amsterdam Neuroscience - Cellular & Molecular Mechanisms, Functional Genomics |
| Rok vydání: | 2020 |
| Předmět: |
Munc18 Proteins
Vesicle-Associated Membrane Protein 2 Vesicle docking Membrane fusion Neuronal Excitability Neurotransmission Synaptic Transmission Animals Receptor chemistry.chemical_classification Crosslinking VAMP2 General Neuroscience Lipid bilayer fusion General Medicine Rats Amino acid chemistry SNARE Munc18-1 Helix Biophysics sense organs SNARE Proteins Research Article: New Research Protein Binding |
| Zdroj: | André, T, Classen, J, Brenner, P, Betts, M J, Dörr, B, Kreye, S, Zuidinga, B, Meijer, M, Russell, R B, Verhage, M & Söllner, T H 2020, ' The interaction of Munc18-1 Helix 11 and 12 with the central region of the VAMP2 SNARE motif is essential for SNARE templating and synaptic transmission ', eNeuro, vol. 7, no. 6, ENEURO.0278-20.2020, pp. 1-5 . https://doi.org/10.1523/ENEURO.0278-20.2020 eNeuro, 7(6):ENEURO.0278-20.2020, 1-5. Society for Neuroscience eNeuro |
| ISSN: | 2373-2822 |
| DOI: | 10.1523/eneuro.0278-20.2020 |
| Popis: | Sec1/Munc18 proteins play a key role in initiating the assembly of N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complexes, the molecular fusion machinery. Employing comparative structure modeling, site specific crosslinking by single amino acid substitutions with the photoactivatable unnatural amino acid p-Benzoyl-phenylalanine (Bpa) and reconstituted vesicle docking/fusion assays, we mapped the binding interface between Munc18-1 and the neuronal v-SNARE VAMP2 with single amino acid resolution. Our results show that helices 11 and 12 of domain 3a in Munc18-1 interact with the VAMP2 SNARE motif covering the region from layers −4 to +5. Residue Q301 in helix 11 plays a pivotal role in VAMP2 binding and template complex formation. A VAMP2 binding deficient mutant, Munc18-1 Q301D, does not stimulate lipid mixing in a reconstituted fusion assay. The neuronal SNARE-organizer Munc13-1, which also binds VAMP2, does not bypass the requirement for the Munc18-1·VAMP2 interaction. Importantly, Munc18-1 Q301D expression inMunc18-1deficient neurons severely reduces synaptic transmission, demonstrating the physiological significance of the Munc18-1·VAMP2 interaction. |
| Databáze: | OpenAIRE |
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