title: Self-assembly of SNARE fusion proteins into star-shaped oligomers.
abstract: Three evolutionarily conserved proteins known as SNAREs (soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptors) mediate exocytosis from single cell eukaryotes to neurons. Among neuronal SNAREs, syntaxin and SNAP-25 (synaptosome-associated protein of 25 kDa) reside on the plasma membrane, whereas synaptobrevin resides on synaptic vesicles prior to fusion. The SNARE motifs of the three proteins form a helical bundle which probably drives membrane fusion. Since studies in vivo suggested an importance for multiple SNARE complexes in the fusion process, and models appeared in the literature with large numbers of SNARE bundles executing the fusion process, we analysed the quaternary structure of the full-length native SNARE complexes in detail. By employing a preparative immunoaffinity procedure we isolated all of the SNARE complexes from brain, and have shown by size-exclusion chromatography and negative stain electron microscopy that they exist as approx. 30 nm particles containing, most frequently, 3 or 4 bundles emanating from their centre. Using highly purified, individual, full-length SNAREs we demonstrated that the oligomerization of SNAREs into star-shaped particles with 3 to 4 bundles is an intrinsic property of these proteins and is not dependent on other proteins, as previously hypothesized. The average number of the SNARE bundles in the isolated fusion particles corresponds well with the co-operativity observed in calcium-triggered neuronal exocytosis.
authors.affiliation: Medical Research Council, Laboratory of Molecular Biology, MRC Centre, Hills Road, Cambridge CB2 2QH, UK.
authors.forename: Colin
authors.initials: C
authors.lastname: Rickman
authors.forename: Kuang
authors.initials: K
authors.lastname: Hu
authors.forename: Joe
authors.initials: J
authors.lastname: Carroll
authors.forename: Bazbek
authors.initials: B
authors.lastname: Davletov
mesh_terms: D011108:Polymers / Q000737:chemistry*; D020836:Protein Structure, Quaternary
chemical_list: D011108:Polymers