Autoinhibition of Munc18-1 modulates synaptobrevin binding and helps to enable Munc13-dependent regulation of membrane fusion
Munc18-1 orchestrates SNARE complex assembly together with Munc13-1 to mediate neurotransmitter release. Munc18-1 binds to synaptobrevin, but the relevance of this interaction and its relation to Munc13 function are unclear. NMR experiments now show that Munc18-1 binds specifically and non-specifica...
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doaj-ff1c5e9989384ec782f9768b36af6d1a2021-05-05T13:27:24ZengeLife Sciences Publications LtdeLife2050-084X2017-05-01610.7554/eLife.24278Autoinhibition of Munc18-1 modulates synaptobrevin binding and helps to enable Munc13-dependent regulation of membrane fusionEwa Sitarska0Junjie Xu1Seungmee Park2Xiaoxia Liu3Bradley Quade4Karolina Stepien5Kyoko Sugita6Chad A Brautigam7Shuzo Sugita8https://orcid.org/0000-0002-9182-873XJosep Rizo9https://orcid.org/0000-0003-1773-8311Department of Biophysics, University of Texas Southwestern Medical Center, Dallas, United States; Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, United States; Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, United StatesDepartment of Biophysics, University of Texas Southwestern Medical Center, Dallas, United States; Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, United States; Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, United StatesDepartment of Physiology, University of Toronto, Toronto, CanadaDepartment of Biophysics, University of Texas Southwestern Medical Center, Dallas, United States; Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, United States; Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, United StatesDepartment of Biophysics, University of Texas Southwestern Medical Center, Dallas, United States; Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, United States; Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, United StatesDepartment of Biophysics, University of Texas Southwestern Medical Center, Dallas, United States; Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, United States; Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, United StatesDepartment of Physiology, University of Toronto, Toronto, CanadaDepartment of Biophysics, University of Texas Southwestern Medical Center, Dallas, United States; Department of Microbiology, University of Texas Southwestern Medical Center, Dallas, United StatesDepartment of Physiology, University of Toronto, Toronto, CanadaDepartment of Biophysics, University of Texas Southwestern Medical Center, Dallas, United States; Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, United States; Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, United StatesMunc18-1 orchestrates SNARE complex assembly together with Munc13-1 to mediate neurotransmitter release. Munc18-1 binds to synaptobrevin, but the relevance of this interaction and its relation to Munc13 function are unclear. NMR experiments now show that Munc18-1 binds specifically and non-specifically to synaptobrevin. Specific binding is inhibited by a L348R mutation in Munc18-1 and enhanced by a D326K mutation designed to disrupt the ‘furled conformation’ of a Munc18-1 loop. Correspondingly, the activity of Munc18-1 in reconstitution assays that require Munc18-1 and Munc13-1 for membrane fusion is stimulated by the D326K mutation and inhibited by the L348R mutation. Moreover, the D326K mutation allows Munc13-1-independent fusion and leads to a gain-of-function in rescue experiments in Caenorhabditis elegans unc-18 nulls. Together with previous studies, our data support a model whereby Munc18-1 acts as a template for SNARE complex assembly, and autoinhibition of synaptobrevin binding contributes to enabling regulation of neurotransmitter release by Munc13-1.https://elifesciences.org/articles/24278Munc18-1Munc13neurotransmitter releaseautoinhibitionprotein NMR |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Ewa Sitarska Junjie Xu Seungmee Park Xiaoxia Liu Bradley Quade Karolina Stepien Kyoko Sugita Chad A Brautigam Shuzo Sugita Josep Rizo |
spellingShingle |
Ewa Sitarska Junjie Xu Seungmee Park Xiaoxia Liu Bradley Quade Karolina Stepien Kyoko Sugita Chad A Brautigam Shuzo Sugita Josep Rizo Autoinhibition of Munc18-1 modulates synaptobrevin binding and helps to enable Munc13-dependent regulation of membrane fusion eLife Munc18-1 Munc13 neurotransmitter release autoinhibition protein NMR |
author_facet |
Ewa Sitarska Junjie Xu Seungmee Park Xiaoxia Liu Bradley Quade Karolina Stepien Kyoko Sugita Chad A Brautigam Shuzo Sugita Josep Rizo |
author_sort |
Ewa Sitarska |
title |
Autoinhibition of Munc18-1 modulates synaptobrevin binding and helps to enable Munc13-dependent regulation of membrane fusion |
title_short |
Autoinhibition of Munc18-1 modulates synaptobrevin binding and helps to enable Munc13-dependent regulation of membrane fusion |
title_full |
Autoinhibition of Munc18-1 modulates synaptobrevin binding and helps to enable Munc13-dependent regulation of membrane fusion |
title_fullStr |
Autoinhibition of Munc18-1 modulates synaptobrevin binding and helps to enable Munc13-dependent regulation of membrane fusion |
title_full_unstemmed |
Autoinhibition of Munc18-1 modulates synaptobrevin binding and helps to enable Munc13-dependent regulation of membrane fusion |
title_sort |
autoinhibition of munc18-1 modulates synaptobrevin binding and helps to enable munc13-dependent regulation of membrane fusion |
publisher |
eLife Sciences Publications Ltd |
series |
eLife |
issn |
2050-084X |
publishDate |
2017-05-01 |
description |
Munc18-1 orchestrates SNARE complex assembly together with Munc13-1 to mediate neurotransmitter release. Munc18-1 binds to synaptobrevin, but the relevance of this interaction and its relation to Munc13 function are unclear. NMR experiments now show that Munc18-1 binds specifically and non-specifically to synaptobrevin. Specific binding is inhibited by a L348R mutation in Munc18-1 and enhanced by a D326K mutation designed to disrupt the ‘furled conformation’ of a Munc18-1 loop. Correspondingly, the activity of Munc18-1 in reconstitution assays that require Munc18-1 and Munc13-1 for membrane fusion is stimulated by the D326K mutation and inhibited by the L348R mutation. Moreover, the D326K mutation allows Munc13-1-independent fusion and leads to a gain-of-function in rescue experiments in Caenorhabditis elegans unc-18 nulls. Together with previous studies, our data support a model whereby Munc18-1 acts as a template for SNARE complex assembly, and autoinhibition of synaptobrevin binding contributes to enabling regulation of neurotransmitter release by Munc13-1. |
topic |
Munc18-1 Munc13 neurotransmitter release autoinhibition protein NMR |
url |
https://elifesciences.org/articles/24278 |
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