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Synaptotagmin oligomers are necessary and can be sufficient to form a Ca(2+)‐sensitive fusion clamp

The buttressed‐ring hypothesis, supported by recent cryo‐electron tomography analysis of docked synaptic‐like vesicles in neuroendocrine cells, postulates that prefusion SNAREpins are stabilized and organized by Synaptotagmin (Syt) ring‐like oligomers. Here, we use a reconstituted single‐vesicle fus...

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Detalles Bibliográficos
Autores principales: Ramakrishnan, Sathish, Bera, Manindra, Coleman, Jeff, Krishnakumar, Shyam S., Pincet, Frederic, Rothman, James E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6349546/
https://www.ncbi.nlm.nih.gov/pubmed/30570144
http://dx.doi.org/10.1002/1873-3468.13317
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author Ramakrishnan, Sathish
Bera, Manindra
Coleman, Jeff
Krishnakumar, Shyam S.
Pincet, Frederic
Rothman, James E.
author_facet Ramakrishnan, Sathish
Bera, Manindra
Coleman, Jeff
Krishnakumar, Shyam S.
Pincet, Frederic
Rothman, James E.
author_sort Ramakrishnan, Sathish
collection PubMed
description The buttressed‐ring hypothesis, supported by recent cryo‐electron tomography analysis of docked synaptic‐like vesicles in neuroendocrine cells, postulates that prefusion SNAREpins are stabilized and organized by Synaptotagmin (Syt) ring‐like oligomers. Here, we use a reconstituted single‐vesicle fusion analysis to test the prediction that destabilizing the Syt1 oligomers destabilizes the clamp and results in spontaneous fusion in the absence of Ca(2+). Vesicles in which Syt oligomerization is compromised by a ring‐destabilizing mutation dock and diffuse freely on the bilayer until they fuse spontaneously, similar to vesicles containing only v‐SNAREs. In contrast, vesicles containing wild‐type Syt are immobile as soon as they attach to the bilayer and remain frozen in place, up to at least 1 h until fusion is triggered by Ca(2+).
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spelling pubmed-63495462019-08-09 Synaptotagmin oligomers are necessary and can be sufficient to form a Ca(2+)‐sensitive fusion clamp Ramakrishnan, Sathish Bera, Manindra Coleman, Jeff Krishnakumar, Shyam S. Pincet, Frederic Rothman, James E. FEBS Lett Research Letters The buttressed‐ring hypothesis, supported by recent cryo‐electron tomography analysis of docked synaptic‐like vesicles in neuroendocrine cells, postulates that prefusion SNAREpins are stabilized and organized by Synaptotagmin (Syt) ring‐like oligomers. Here, we use a reconstituted single‐vesicle fusion analysis to test the prediction that destabilizing the Syt1 oligomers destabilizes the clamp and results in spontaneous fusion in the absence of Ca(2+). Vesicles in which Syt oligomerization is compromised by a ring‐destabilizing mutation dock and diffuse freely on the bilayer until they fuse spontaneously, similar to vesicles containing only v‐SNAREs. In contrast, vesicles containing wild‐type Syt are immobile as soon as they attach to the bilayer and remain frozen in place, up to at least 1 h until fusion is triggered by Ca(2+). John Wiley and Sons Inc. 2019-01-18 2019-01 /pmc/articles/PMC6349546/ /pubmed/30570144 http://dx.doi.org/10.1002/1873-3468.13317 Text en © 2018 The Authors. FEBS Letters published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies. This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Letters
Ramakrishnan, Sathish
Bera, Manindra
Coleman, Jeff
Krishnakumar, Shyam S.
Pincet, Frederic
Rothman, James E.
Synaptotagmin oligomers are necessary and can be sufficient to form a Ca(2+)‐sensitive fusion clamp
title Synaptotagmin oligomers are necessary and can be sufficient to form a Ca(2+)‐sensitive fusion clamp
title_full Synaptotagmin oligomers are necessary and can be sufficient to form a Ca(2+)‐sensitive fusion clamp
title_fullStr Synaptotagmin oligomers are necessary and can be sufficient to form a Ca(2+)‐sensitive fusion clamp
title_full_unstemmed Synaptotagmin oligomers are necessary and can be sufficient to form a Ca(2+)‐sensitive fusion clamp
title_short Synaptotagmin oligomers are necessary and can be sufficient to form a Ca(2+)‐sensitive fusion clamp
title_sort synaptotagmin oligomers are necessary and can be sufficient to form a ca(2+)‐sensitive fusion clamp
topic Research Letters
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6349546/
https://www.ncbi.nlm.nih.gov/pubmed/30570144
http://dx.doi.org/10.1002/1873-3468.13317
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