Disassembly of the reconstituted synaptic vesicle membrane fusion complex in vitro

Tetsuya Hayashi, S. Yamasaki, S. Nauenburg, T. Binz, H. Niemann

Research output: Contribution to journalArticle

221 Citations (Scopus)

Abstract

The interaction of the presynaptic membrane proteins SNAP-25 and syntaxin with the synaptic vesicle protein synaptobrevin (VAMP) plays a key role in the regulated exocytosis of neurotransmitters. Clostridial neurotoxins, which proteolyze these polypeptides, are potent inhibitors of neurotransmission. The cytoplasmic domains of the three membrane proteins join into a tight SDS-resistant complex. Here, we show that this reconstituted complex, as well as heterodimers composed of syntaxin and SNAP-25, can be disassembled by the concerted action of the N-ethylmaleimide-sensitive factor, NSF, and the soluble NSF attachment protein, α-SNAP α-SNAP binds to predicted α-helical coiled-coil regions of syntaxin and SNAP-25, shown previously to be engaged in their direct interaction. Synaptobrevin, although incapable of binding α-SNAP individually, induced a third α-SNAP binding site when associated with syntaxin and SNAP-25 into heterotrimers. NSF released prebound α-SNAP from full-length syntaxin but not from a syntaxin derivative truncated at the N-terminus. Disassembly of complexes containing this syntaxin mutant was impaired, indicating a critical role for the N-terminal domain in the α-SNAP/NSF-mediated dissociation process. Complexes containing C-terminally deleted SNAP-25 derivatives, as generated by botulinal toxins type A and E, were dissociated more efficiently. In contrast, the N-terminal fragment generated from synaptobrevin by botulinal toxin type F produced an SDS-sensitive complex that was poorly dissociated.

Original languageEnglish
Pages (from-to)2317-2325
Number of pages9
JournalEMBO Journal
Volume14
Issue number10
Publication statusPublished - Jan 1 1995
Externally publishedYes

Fingerprint

Qa-SNARE Proteins
Synaptic Membranes
Membrane Fusion
Synaptic Vesicles
Fusion reactions
Membranes
R-SNARE Proteins
Botulinum Toxins
Clostridium botulinum type F
Clostridium botulinum type E
Membrane Proteins
Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins
N-Ethylmaleimide-Sensitive Proteins
Derivatives
Tetanus Toxin
Exocytosis
In Vitro Techniques
Synaptic Transmission
Neurotransmitter Agents
Binding Sites

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)
  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Hayashi, T., Yamasaki, S., Nauenburg, S., Binz, T., & Niemann, H. (1995). Disassembly of the reconstituted synaptic vesicle membrane fusion complex in vitro. EMBO Journal, 14(10), 2317-2325.

Disassembly of the reconstituted synaptic vesicle membrane fusion complex in vitro. / Hayashi, Tetsuya; Yamasaki, S.; Nauenburg, S.; Binz, T.; Niemann, H.

In: EMBO Journal, Vol. 14, No. 10, 01.01.1995, p. 2317-2325.

Research output: Contribution to journalArticle

Hayashi, T, Yamasaki, S, Nauenburg, S, Binz, T & Niemann, H 1995, 'Disassembly of the reconstituted synaptic vesicle membrane fusion complex in vitro', EMBO Journal, vol. 14, no. 10, pp. 2317-2325.
Hayashi T, Yamasaki S, Nauenburg S, Binz T, Niemann H. Disassembly of the reconstituted synaptic vesicle membrane fusion complex in vitro. EMBO Journal. 1995 Jan 1;14(10):2317-2325.
Hayashi, Tetsuya ; Yamasaki, S. ; Nauenburg, S. ; Binz, T. ; Niemann, H. / Disassembly of the reconstituted synaptic vesicle membrane fusion complex in vitro. In: EMBO Journal. 1995 ; Vol. 14, No. 10. pp. 2317-2325.
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