Binding of the Munc13-1 MUN domain to membrane-anchored SNARE complexes

Rong Guan, Han Dai, Jose Rizo-Rey

Research output: Contribution to journalArticle

76 Citations (Scopus)

Abstract

The core of the membrane fusion machinery that governs neurotransmitter release includes the SNARE proteins syntaxin-1, SNAP-25 and synaptobrevin, which form a tight "SNARE complex", and Munc18-1, which binds to the SNARE complex and to syntaxin-1 folded into a closed conformation. Release is also controlled by specialized proteins such as complexins, which also bind to the SNARE complex, and unc13/Munc13s, which are crucial for synaptic vesicle priming and were proposed to open syntaxin-1, promoting SNARE complex assembly. However, the biochemical basis for unc13/Munc13 function and its relationship to other SNARE interactions are unclear. To address this question, we have analyzed interactions of the MUN domain of Munc13-1, which is key for this priming function, using solution binding assays and cofloatation experiments with SNARE-containing proteoliposomes. Our results indicate that the Munc13-1 MUN domain binds to membrane-anchored SNARE complexes, even though binding is barely detectable in solution. The MUN domain appears to compete with Munc18-1 but not with complexin-1 for SNARE complex binding, although more quantitative assays will be required to verify these conclusions. Moreover, our data also uncover interactions of membrane-anchored syntaxin-1/SNAP-25 heterodimers with the MUN domain, Munc18-1 and complexin-1. The interaction with complexin-1 is surprising, as it was not observed in previous solution studies. Our results emphasize the importance of studying interactions within the neurotransmitter release machinery in a native membrane environment, and suggest that unc13/Munc13s may provide a template to assemble syntaxin-1/SNAP-25 heterodimers, leading to an acceptor complex for synaptobrevin.

Original languageEnglish (US)
Pages (from-to)1474-1481
Number of pages8
JournalBiochemistry
Volume47
Issue number6
DOIs
StatePublished - Feb 12 2008

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SNARE Proteins
Syntaxin 1
Membranes
R-SNARE Proteins
Machinery
Neurotransmitter Agents
Assays
Membrane Fusion
Synaptic Vesicles
Conformations
Fusion reactions

ASJC Scopus subject areas

  • Biochemistry

Cite this

Binding of the Munc13-1 MUN domain to membrane-anchored SNARE complexes. / Guan, Rong; Dai, Han; Rizo-Rey, Jose.

In: Biochemistry, Vol. 47, No. 6, 12.02.2008, p. 1474-1481.

Research output: Contribution to journalArticle

Guan, Rong ; Dai, Han ; Rizo-Rey, Jose. / Binding of the Munc13-1 MUN domain to membrane-anchored SNARE complexes. In: Biochemistry. 2008 ; Vol. 47, No. 6. pp. 1474-1481.
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