title: Allosteric control of syntaxin 1a by Munc18-1: characterization of the open and closed conformations of syntaxin.
abstract: Syntaxin 1a is a plasma membrane soluble N-ethylmaleimide-sensitive factor attachment receptor protein (SNARE) that contains an H3 domain (SNARE motif) and a regulatory Habc domain. These regions associate to produce a closed state, which is generally thought to suppress assembly of syntaxin into the SNARE complex. However, the molecular nature of the closed and open states of syntaxin is not well defined. Here, we use electron paramagnetic resonance spectroscopy to characterize conformational exchange in syntaxin. The data indicate that the H3 segment is in equilibrium between ordered and disordered states that have significant populations. In solution, the central region of the H3 segment is positioned close to the Habc domain and the configuration of syntaxin 1a is dominated by a closed state. However, an open state is enhanced in full-length membrane reconstituted syntaxin. Munc18-1 binding alters the equilibrium along H3 to favor the ordered, folded state. Munc18 also suppresses the minor open population and narrows the distance distributions between H3 and Habc. The allosteric control exhibited by Munc18 on the H3 segment and the suppression of the minor open component may both play a role in regulating membrane fusion by controlling the assembly of syntaxin into the SNARE complex.
authors.affiliation: Department of Chemistry, University of Virginia, Charlottesville, Virginia, USA.
authors.forename: Damian
authors.initials: D
authors.lastname: Dawidowski
authors.forename: David S
authors.initials: DS
authors.lastname: Cafiso
mesh_terms: D000494:Allosteric Regulation; D002462:Cell Membrane / Q000378:metabolism; D004578:Electron Spin Resonance Spectroscopy; D008958:Models, Molecular; D051938:Munc18 Proteins / Q000378:metabolism*; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D013113:Spin Labels; D050827:Syntaxin 1 / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D051938:Munc18 Proteins; D013113:Spin Labels; D050827:Syntaxin 1
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