A protein assembly-disassembly pathway in vitro that may correspond to sequential steps of synaptic vesicle docking, activation, and fusion

Thomas Söllner; Mark K. Bennett; Sidney W. Whiteheart; Richard H. Scheller; James E. Rothman

doi:10.1016/0092-8674(93)90376-2

A protein assembly-disassembly pathway in vitro that may correspond to sequential steps of synaptic vesicle docking, activation, and fusion

Thomas Söllner, Mark K. Bennett, Sidney W. Whiteheart, Richard H. Scheller, James E. Rothman

Molecular and Cellular Biochemistry

Research output: Contribution to journal › Article › peer-review

1640 Scopus citations

Abstract

The SNARE hypothesis holds that a transport vesicle chooses its target for fusion when a soluble NSF attachment protein (SNAP) receptor on the vesicle (v-SNARE) pairs with its cognate t-SNARE at the target membrane. Three synaptosomal membrane proteins have previously been identified: syntaxin, SNAP-25 (t-SNAREs), and vesicle-associated membrane protein (VAMP) (v-SNARE); all assemble with SNAPs and NSF into 20S fusion particles. We now report that in the absence of SNAP and NSF, these three SNAREs form a stable complex that can also bind synaptotagmin. Synaptotagmin is displaced by α-SNAP, suggesting that these two proteins share binding sites on the SNARE complex and implying that synaptotagmin operates as a "clamp" to prevent fusion from proceeding in the absence of a signal. The α-SNAP-SNARE complex can bind NSF, and NSF-dependent hydrolysis of ATP dissociates the complex, separating syntaxin, SNAP-25, and VAMP. ATP hydrolysis by NSF may provide motion to initiate bilayer fusion.

Original language	English
Pages (from-to)	409-418
Number of pages	10
Journal	Cell
Volume	75
Issue number	3
DOIs	https://doi.org/10.1016/0092-8674(93)90376-2
State	Published - Nov 5 1993

Bibliographical note

Funding Information:
Correspondence should be addressed to J. E. Ft. We thank Masami Nagahama for construction of the His+,-NSF-Myc clone, Willa Bellamy for help with the manuscript, and Wayne Patton for gel scans and analysis. This work was supported by a National Institutes of Health grant to J. E. R., by the Matherscharitable Foundation and a National Institute of Mental Health grant to R. Ii. S., by a fellowship of the Deutsche Forschungsgemeinschaft to T. S., and by a fellowship from the Jane Coffin Childs Memorial Fund for Medical Research to s. w. w.

ASJC Scopus subject areas

Biochemistry, Genetics and Molecular Biology (all)

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10.1016/0092-8674(93)90376-2

Cite this

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title = "A protein assembly-disassembly pathway in vitro that may correspond to sequential steps of synaptic vesicle docking, activation, and fusion",

abstract = "The SNARE hypothesis holds that a transport vesicle chooses its target for fusion when a soluble NSF attachment protein (SNAP) receptor on the vesicle (v-SNARE) pairs with its cognate t-SNARE at the target membrane. Three synaptosomal membrane proteins have previously been identified: syntaxin, SNAP-25 (t-SNAREs), and vesicle-associated membrane protein (VAMP) (v-SNARE); all assemble with SNAPs and NSF into 20S fusion particles. We now report that in the absence of SNAP and NSF, these three SNAREs form a stable complex that can also bind synaptotagmin. Synaptotagmin is displaced by α-SNAP, suggesting that these two proteins share binding sites on the SNARE complex and implying that synaptotagmin operates as a {"}clamp{"} to prevent fusion from proceeding in the absence of a signal. The α-SNAP-SNARE complex can bind NSF, and NSF-dependent hydrolysis of ATP dissociates the complex, separating syntaxin, SNAP-25, and VAMP. ATP hydrolysis by NSF may provide motion to initiate bilayer fusion.",

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note = "Funding Information: Correspondence should be addressed to J. E. Ft. We thank Masami Nagahama for construction of the His+,-NSF-Myc clone, Willa Bellamy for help with the manuscript, and Wayne Patton for gel scans and analysis. This work was supported by a National Institutes of Health grant to J. E. R., by the Matherscharitable Foundation and a National Institute of Mental Health grant to R. Ii. S., by a fellowship of the Deutsche Forschungsgemeinschaft to T. S., and by a fellowship from the Jane Coffin Childs Memorial Fund for Medical Research to s. w. w.",

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AU - Rothman, James E.

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N2 - The SNARE hypothesis holds that a transport vesicle chooses its target for fusion when a soluble NSF attachment protein (SNAP) receptor on the vesicle (v-SNARE) pairs with its cognate t-SNARE at the target membrane. Three synaptosomal membrane proteins have previously been identified: syntaxin, SNAP-25 (t-SNAREs), and vesicle-associated membrane protein (VAMP) (v-SNARE); all assemble with SNAPs and NSF into 20S fusion particles. We now report that in the absence of SNAP and NSF, these three SNAREs form a stable complex that can also bind synaptotagmin. Synaptotagmin is displaced by α-SNAP, suggesting that these two proteins share binding sites on the SNARE complex and implying that synaptotagmin operates as a "clamp" to prevent fusion from proceeding in the absence of a signal. The α-SNAP-SNARE complex can bind NSF, and NSF-dependent hydrolysis of ATP dissociates the complex, separating syntaxin, SNAP-25, and VAMP. ATP hydrolysis by NSF may provide motion to initiate bilayer fusion.

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A protein assembly-disassembly pathway in vitro that may correspond to sequential steps of synaptic vesicle docking, activation, and fusion

Abstract

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