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Conformational change of Syntaxin-3b in regulating SNARE complex assembly in the ribbon synapses
Neurotransmitter release of synaptic vesicles relies on the assembly of the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complex, consisting of syntaxin and SNAP-25 on the plasma membrane and synaptobrevin on the synaptic vesicle. The formation of the SNARE complex p...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9166750/ https://www.ncbi.nlm.nih.gov/pubmed/35661757 http://dx.doi.org/10.1038/s41598-022-09654-3 |
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author | Gething, Claire Ferrar, Joshua Misra, Bishal Howells, Giovanni Andrzejewski, Alexa L. Bowen, Mark E. Choi, Ucheor B. |
author_facet | Gething, Claire Ferrar, Joshua Misra, Bishal Howells, Giovanni Andrzejewski, Alexa L. Bowen, Mark E. Choi, Ucheor B. |
author_sort | Gething, Claire |
collection | PubMed |
description | Neurotransmitter release of synaptic vesicles relies on the assembly of the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complex, consisting of syntaxin and SNAP-25 on the plasma membrane and synaptobrevin on the synaptic vesicle. The formation of the SNARE complex progressively zippers towards the membranes, which drives membrane fusion between the plasma membrane and the synaptic vesicle. However, the underlying molecular mechanism of SNARE complex regulation is unclear. In this study, we investigated the syntaxin-3b isoform found in the retinal ribbon synapses using single-molecule fluorescence resonance energy transfer (smFRET) to monitor the conformational changes of syntaxin-3b that modulate the SNARE complex formation. We found that syntaxin-3b is predominantly in a self-inhibiting closed conformation, inefficiently forming the ternary SNARE complex. Conversely, a phosphomimetic mutation (T14E) at the N-terminal region of syntaxin-3b promoted the open conformation, similar to the constitutively open form of syntaxin LE mutant. When syntaxin-3b is bound to Munc18-1, SNARE complex formation is almost completely blocked. Surprisingly, the T14E mutation of syntaxin-3b partially abolishes Munc18-1 regulation, acting as a conformational switch to trigger SNARE complex assembly. Thus, we suggest a model where the conformational change of syntaxin-3b induced by phosphorylation initiates the release of neurotransmitters in the ribbon synapses. |
format | Online Article Text |
id | pubmed-9166750 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-91667502022-06-05 Conformational change of Syntaxin-3b in regulating SNARE complex assembly in the ribbon synapses Gething, Claire Ferrar, Joshua Misra, Bishal Howells, Giovanni Andrzejewski, Alexa L. Bowen, Mark E. Choi, Ucheor B. Sci Rep Article Neurotransmitter release of synaptic vesicles relies on the assembly of the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complex, consisting of syntaxin and SNAP-25 on the plasma membrane and synaptobrevin on the synaptic vesicle. The formation of the SNARE complex progressively zippers towards the membranes, which drives membrane fusion between the plasma membrane and the synaptic vesicle. However, the underlying molecular mechanism of SNARE complex regulation is unclear. In this study, we investigated the syntaxin-3b isoform found in the retinal ribbon synapses using single-molecule fluorescence resonance energy transfer (smFRET) to monitor the conformational changes of syntaxin-3b that modulate the SNARE complex formation. We found that syntaxin-3b is predominantly in a self-inhibiting closed conformation, inefficiently forming the ternary SNARE complex. Conversely, a phosphomimetic mutation (T14E) at the N-terminal region of syntaxin-3b promoted the open conformation, similar to the constitutively open form of syntaxin LE mutant. When syntaxin-3b is bound to Munc18-1, SNARE complex formation is almost completely blocked. Surprisingly, the T14E mutation of syntaxin-3b partially abolishes Munc18-1 regulation, acting as a conformational switch to trigger SNARE complex assembly. Thus, we suggest a model where the conformational change of syntaxin-3b induced by phosphorylation initiates the release of neurotransmitters in the ribbon synapses. Nature Publishing Group UK 2022-06-03 /pmc/articles/PMC9166750/ /pubmed/35661757 http://dx.doi.org/10.1038/s41598-022-09654-3 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Gething, Claire Ferrar, Joshua Misra, Bishal Howells, Giovanni Andrzejewski, Alexa L. Bowen, Mark E. Choi, Ucheor B. Conformational change of Syntaxin-3b in regulating SNARE complex assembly in the ribbon synapses |
title | Conformational change of Syntaxin-3b in regulating SNARE complex assembly in the ribbon synapses |
title_full | Conformational change of Syntaxin-3b in regulating SNARE complex assembly in the ribbon synapses |
title_fullStr | Conformational change of Syntaxin-3b in regulating SNARE complex assembly in the ribbon synapses |
title_full_unstemmed | Conformational change of Syntaxin-3b in regulating SNARE complex assembly in the ribbon synapses |
title_short | Conformational change of Syntaxin-3b in regulating SNARE complex assembly in the ribbon synapses |
title_sort | conformational change of syntaxin-3b in regulating snare complex assembly in the ribbon synapses |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9166750/ https://www.ncbi.nlm.nih.gov/pubmed/35661757 http://dx.doi.org/10.1038/s41598-022-09654-3 |
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