title: Munc18a controls SNARE assembly through its interaction with the syntaxin N-peptide.
abstract: Sec1/Munc18-like (SM) proteins functionally interact with SNARE proteins in vesicular fusion. Despite their high sequence conservation, structurally disparate binding modes for SM proteins with syntaxins have been observed. Several SM proteins appear to bind only to a short peptide present at the N terminus of syntaxin, designated the N-peptide, while Munc18a binds to a 'closed' conformation formed by the remaining portion of syntaxin 1a. Here, we show that the syntaxin 16 N-peptide binds to the SM protein Vps45, but the remainder of syntaxin 16 strongly enhances the affinity of the interaction. Likewise, the N-peptide of syntaxin 1a serves as a second binding site in the Munc18a/syntaxin 1a complex. When the syntaxin 1a N-peptide is bound to Munc18a, SNARE complex formation is blocked. Removal of the N-peptide enables binding of syntaxin 1a to its partner SNARE SNAP-25, while still bound to Munc18a. This suggests that Munc18a controls the accessibility of syntaxin 1a to its partners, a role that might be common to all SM proteins.
authors.affiliation: Research Group Structural Biochemistry, Department of Neurobiology, Max-Planck-Institute for Biophysical Chemistry, Göttingen, Germany.
authors.forename: Pawel
authors.initials: P
authors.lastname: Burkhardt
authors.forename: Douglas A
authors.initials: DA
authors.lastname: Hattendorf
authors.forename: William I
authors.initials: WI
authors.lastname: Weis
authors.forename: Dirk
authors.initials: D
authors.lastname: Fasshauer
mesh_terms: D000818:Animals; D001665:Binding Sites; D002151:Calorimetry; D018360:Crystallography, X-Ray; D008565:Membrane Proteins / Q000378:metabolism; D008958:Models, Molecular; D051938:Munc18 Proteins / Q000378:metabolism*; D050505:Mutant Proteins / Q000378:metabolism; D010455:Peptides / Q000378:metabolism*; D011485:Protein Binding; D017433:Protein Structure, Secondary; D051381:Rats; D050600:SNARE Proteins / Q000378:metabolism*; D050827:Syntaxin 1 / Q000737:chemistry* / Q000378:metabolism*; D013816:Thermodynamics; D033921:Vesicular Transport Proteins / Q000378:metabolism
chemical_list: D008565:Membrane Proteins; D051938:Munc18 Proteins; D050505:Mutant Proteins; D010455:Peptides; D050600:SNARE Proteins; D050827:Syntaxin 1; D033921:Vesicular Transport Proteins; C498420:Vps45 protein, rat