Self-association of the H3 Region of Syntaxin 1A

Misura, Kira M. S.; Scheller, Richard H.; Weis, William I.

doi:10.1074/jbc.m009636200

Cited by 65 publications

(28 citation statements)

References 36 publications

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“…2D). CAPS also failed to interact with the soluble cytoplasmic domain syntaxin-(1-264) likely because it is in a "closed" conformation that occludes interactions with the H3 domain (32). The SPR results confirmed that CAPS interacts with the H3-linker domain of syntaxin-1.…”

Section: Caps-1 Binds Membrane-integrated Syntaxin-1-caps-1mentioning

confidence: 66%

Novel Interactions of CAPS (Ca2+-dependent Activator Protein for Secretion) with the Three Neuronal SNARE Proteins Required for Vesicle Fusion

Daily

Boswell

James

et al. 2010

Journal of Biological Chemistry

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CAPS (aka CADPS)Peptide and neurotransmitter release from endocrine cells and neurons occurs by the regulated exocytic fusion of secretory vesicles with the plasma membrane (1). Membrane fusion is mediated by trans complexes of SNARE proteins that bridge the vesicle and plasma membrane to promote close membrane apposition and bilayer mixing (2). The SNARE complexes for endocrine and neuronal vesicle fusion consist of parallel bundles of four ␣-helical SNARE motifs with one helix contributed by the vesicle v-SNARE vesicle-associated membrane protein 2 (VAMP-2) 2 (aka synaptobrevin), one helix contributed by the plasma membrane t-SNARE syntaxin-1, and two helices contributed by the plasma membrane t-SNARE SNAP-25 (3). Based on a central layer residue (arginine or glutamine) and helix position in the SNARE bundle, these proteins are also classified as R-, Qa-, and QbQc-SNAREs, respectively (4). In vitro studies suggested a "zipper" model for the assembly of ternary SNARE complexes involving the pairing of syntaxin-1 (Qa) with SNAP-25 (QbQc) followed by the N-to C-terminal insertion of the R-SNARE VAMP-2 (5, 6).

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Section: Caps-1 Binds Membrane-integrated Syntaxin-1-caps-1mentioning

confidence: 66%

Novel Interactions of CAPS (Ca2+-dependent Activator Protein for Secretion) with the Three Neuronal SNARE Proteins Required for Vesicle Fusion

Daily

Boswell

James

et al. 2010

Journal of Biological Chemistry

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“…Like the SNAP-23N homotetramer, the neuronal heterotetramer and complexes between the SNAP-25 coiled-coil domains with the syntaxin-1a coiled-coil domain are arranged as parallel four-helix bundles (7,(23)(24)(25). In contrast, a syntaxin-1a homotetramer contains two pairs of helices that run antiparallel to one another (26). The neuronal heterotetrameric complex has a much larger left-handed helical twist compared with the SNAP-23N homotetramer, because each chain in the heterotetramer has a slightly different conformation.…”

Section: Fig 2 Snare Peptide Binding Assay a Individual Snare Pepmentioning

confidence: 99%

“…Each intermolecular layer of our structure has the same four hydrophobic side chains as a result of the 4-fold symmetry, whereas other SNARE complex structures have different hydrophobic side chains in each layer. The latter is also true for the syntaxin-1a homotetrameric structure, because the chains in this complex are antiparallel (26). Bulky hydrophobic side chains are accommodated by complementary smaller hydrophobic ones in each layer of these other SNARE complex structures (4).…”

Section: Fig 2 Snare Peptide Binding Assay a Individual Snare Pepmentioning

confidence: 99%

Homotetrameric Structure of the SNAP-23 N-terminal Coiled-coil Domain

Freedman

Song

et al. 2003

Journal of Biological Chemistry

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SNARE proteins mediate intracellular membrane fusion by forming a coiled-coil complex to merge opposing membranes. A "fusion-active" neuronal SNARE complex is a parallel four-helix bundle containing two coiled-coil domains from SNAP-25 and one coiled-coil domain each from syntaxin-1a and VAMP-2. "Prefusion" assembly intermediate complexes can also form from these SNAREs. We studied the N-terminal coiled-coil domain of SNAP-23 (SNAP-23N), a non-neuronal homologue of SNAP-25, and its interaction with other coiled-coil domains. SNAP-23N can assemble spontaneously with the coiled-coil domains from SNAP-23C, syntaxin-4, and VAMP-3 to form a heterotetrameric complex. Unexpectedly, pure SNAP-23N crystallizes as a coiled-coil homotetrameric complex. The four helices have a parallel orientation and are symmetrical about the long axis. The complex is stabilized through the interaction of conserved hydrophobic residues comprising the a and d positions of the coiled-coil heptad repeats. In addition, a central, highly conserved glutamine residue (Gln-48) is buried within the interface by hydrogen bonding between glutamine side chains derived from adjacent subunits and to solvent molecules. A comparison of the SNAP-23N structure to other SNARE complex structures reveals how a simple coiled-coil motif can form diverse SNARE complexes.Endosomal trafficking and vesicle secretion in eukaryotic cells requires fusion of membrane compartments. Synaptic vesicle secretion in neurons has served as a paradigm for understanding the molecular basis for membrane fusion (1). In general, a v-SNARE 1 called VAMP (synaptobrevin) is associated with the vesicle membrane and two t-SNAREs called SNAP-25 and syntaxin are associated with the plasma or target membrane. According to the SNARE hypothesis, specific pairing between v-and t-SNAREs brings together two membranes for fusion (2). Following fusion, NSF and SNAPs dissociate the SNARE complex in an ATP-dependent reaction (3). SNARE proteins contain coiled-coil regions that mediate complex formation (4). VAMP and syntaxin proteins each have a single SNARE-interacting coiled-coil domain followed by a C-terminal transmembrane domain, whereas SNAP-25 proteins have N-and C-terminal coiled-coil domains separated by a loop that attaches to membranes via palmitoylated cysteines (5). The coiled-coil sequences are highly conserved among SNARE families (4). The homologous regions are made up of ϳ8 heptad repeats in which the hydrophobic residues in the a and d positions of each repeat align in 16 layers (4). The central "0" layer is uniquely polar and consists of a glutamine in syntaxin and the SNAP-25 N-and C-terminal coiled-coil domains and an arginine in the VAMP coiled-coil domain. The structure of a core SNARE complex from neurons revealed a parallel four-helix bundle composed of two coiled-coil domains from SNAP-25, and one coiled-coil domain each from syntaxin-1a and VAMP-2 (6, 7). The parallel orientation of SNARE proteins attached to opposite membranes suggests a "zipperlike" mechanism for membran...

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“…In addition to the well studied heterotrimeric neuronal and yeast plasma membrane SNARE complexes, other homo-and heteromeric interactions between individual SNAREs have been found and characterized (26)(27)(28). These alternate complexes are less stable than the heterotrimeric SNARE complex, but the relevance of these interactions is unknown.…”

mentioning

confidence: 99%

Single-molecule studies of SNARE complex assembly reveal parallel and antiparallel configurations

Weninger

Bowen

Chu

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

122

152

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Vesicle fusion in eukaryotes is thought to involve the assembly of a highly conserved family of proteins termed soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) into a highly stable parallel four-helix bundle. We have used intermolecular single-molecule fluorescence resonance energy transfer to characterize preassembled neuronal SNARE complexes consisting of syntaxin, synaptobrevin, and synaptosome-associated protein of 25 kDa on deposited lipid bilayers. Surprisingly, we found a mixture of parallel as well as antiparallel configurations involving the SNARE motifs of syntaxin and synaptobrevin as well as those of syntaxin and synaptosome-associated protein of 25 kDa. The subpopulation with the parallel four-helix bundle configuration could be greatly enriched by an additional purification step in the presence of denaturant, indicating that the parallel configuration is the energetically most favorable state. Interconversion between the configurations was not observed. From this observation, we infer the conversion rate to be <1.5 h ؊1 . The existence of antiparallel configurations suggests a regulatory role of chaperones, such as N-ethylmaleimide-sensitive factor, or the membrane environment during SNARE complex assembly in vivo, and it could be a partial explanation for the relatively slow rates of vesicle fusion observed by reconstituted fusion experiments in vitro.A ll membrane fusion events in eukaryotes are thought to involve a highly conserved family of proteins termed soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) (1, 2). The neuronal SNAREs syntaxin, synaptosome-associated protein of 25 kDa (SNAP-25), and synaptobrevin are involved in the Ca 2ϩ -dependent fusion of synaptic vesicles with the presynaptic plasma membrane (3). Before synaptic vesicle docking, the individual SNAREs, syntaxin and SNAP-25, are primarily found on the plasma membrane, where they may exist as unfolded monomers, preformed binary complexes, or in complex with chaperones. At some point during docking and fusion of synaptic vesicles, heterotrimeric SNARE complexes form between syntaxin, SNAP-25, and the synaptic vesicle SNARE, synaptobrevin. SNARE complex formation is thought to occur first in trans with proteins on opposite membranes, and end with formation of a cis complex with proteins residing in the same membrane. Thus, SNARE complex crystal structures (4-7) probably represent the state of the SNARE complex at the endpoint of the fusion reaction (8).The inhibition of neurotransmitter release after the specific cleavage of any one of the SNAREs by clostridial neurotoxin proteases supports the fundamental role of the SNARE proteins in synaptic vesicle fusion (9, 10). However, whereas high concentrations of SNAREs have been shown to be sufficient to fuse synthetic liposomes in vitro (11), the molecular relationship between SNARE complex formation and Ca 2ϩ -triggered vesicle-membrane fusion in vivo remains unclear (12).Syntaxin, SNAP-25, and synaptobrevin have been the...

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Self-association of the H3 Region of Syntaxin 1A

Cited by 65 publications

References 36 publications

Novel Interactions of CAPS (Ca2+-dependent Activator Protein for Secretion) with the Three Neuronal SNARE Proteins Required for Vesicle Fusion

Novel Interactions of CAPS (Ca2+-dependent Activator Protein for Secretion) with the Three Neuronal SNARE Proteins Required for Vesicle Fusion

Homotetrameric Structure of the SNAP-23 N-terminal Coiled-coil Domain

Single-molecule studies of SNARE complex assembly reveal parallel and antiparallel configurations

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