Syntaxin 1A Drives Fusion of Large Dense-Core Neurosecretory Granules Into a Planar Lipid Bilayer
dc.contributor.author | McNally, James M. | |
dc.contributor.author | Woodbury, Dixon J. | |
dc.contributor.author | Lemos, Jose R. | |
dc.date | 2022-08-11T08:09:01.000 | |
dc.date.accessioned | 2022-08-23T16:15:48Z | |
dc.date.available | 2022-08-23T16:15:48Z | |
dc.date.issued | 2004-09-17 | |
dc.date.submitted | 2008-11-21 | |
dc.identifier.citation | Cell Biochem Biophys. 2004;41(1):11-24. <a href="http://dx.doi.org/10.1385/CBB:41:1:011 ">Link to article on publisher's site</a> | |
dc.identifier.issn | 1085-9195 (Print) | |
dc.identifier.doi | 10.1385/CBB:41:1:011 | |
dc.identifier.pmid | 15371637 | |
dc.identifier.uri | http://hdl.handle.net/20.500.14038/34179 | |
dc.description.abstract | The SNARE complex, involved in vesicular trafficking and exocytosis, is composed of proteins in the vesicular membrane (v-SNAREs) that intertwine with proteins of the target membrane (t-SNAREs). Our results show that modified large dense-core neurosecretory granules (NSGs), isolated from the bovine neurohypophysis, spontaneously fuse with a planar lipid membrane containing only the t-SNARE syntaxin 1A. This provides evidence that syntaxin alone is able to form a functional fusion complex with native v-SNAREs of the NSG. The fusion was similar to constitutive, not regulated, exocytosis because changes in free [Ca2+] had no effect on the syntaxin-mediated fusion. Several deletion mutants of syntaxin 1A were also tested. The removal of the regulatory domain did not significantly reduce spontaneous fusion. However, a syntaxin deletion mutant consisting of only the transmembrane domain was incapable of eliciting spontaneous fusion. Finally, a soluble form of syntaxin 1A (lacking its transmembrane domain) was used to saturate the free syntaxin-binding sites of modified NSGs. This treatment blocks spontaneous fusion of these granules to a bilayer containing full-length syntaxin 1A. This method provides an effective model system to study possible regulatory components affecting vesicle fusion. | |
dc.language.iso | en_US | |
dc.relation | <a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=15371637&dopt=Abstract">Link to article in PubMed</a> | |
dc.relation.url | http://dx.doi.org/10.1385/CBB:41:1:011 | |
dc.subject | Lipid Bilayers; Secretory Vesicles; Syntaxin 1 | |
dc.subject | Life Sciences | |
dc.subject | Medicine and Health Sciences | |
dc.subject | Neuroscience and Neurobiology | |
dc.title | Syntaxin 1A Drives Fusion of Large Dense-Core Neurosecretory Granules Into a Planar Lipid Bilayer | |
dc.type | Journal Article | |
dc.source.journaltitle | Cell biochemistry and biophysics | |
dc.source.volume | 41 | |
dc.source.issue | 1 | |
dc.identifier.legacycoverpage | https://escholarship.umassmed.edu/gsbs_sp/838 | |
dc.identifier.contextkey | 670494 | |
html.description.abstract | <p>The SNARE complex, involved in vesicular trafficking and exocytosis, is composed of proteins in the vesicular membrane (v-SNAREs) that intertwine with proteins of the target membrane (t-SNAREs). Our results show that modified large dense-core neurosecretory granules (NSGs), isolated from the bovine neurohypophysis, spontaneously fuse with a planar lipid membrane containing only the t-SNARE syntaxin 1A. This provides evidence that syntaxin alone is able to form a functional fusion complex with native v-SNAREs of the NSG. The fusion was similar to constitutive, not regulated, exocytosis because changes in free [Ca2+] had no effect on the syntaxin-mediated fusion. Several deletion mutants of syntaxin 1A were also tested. The removal of the regulatory domain did not significantly reduce spontaneous fusion. However, a syntaxin deletion mutant consisting of only the transmembrane domain was incapable of eliciting spontaneous fusion. Finally, a soluble form of syntaxin 1A (lacking its transmembrane domain) was used to saturate the free syntaxin-binding sites of modified NSGs. This treatment blocks spontaneous fusion of these granules to a bilayer containing full-length syntaxin 1A. This method provides an effective model system to study possible regulatory components affecting vesicle fusion.</p> | |
dc.identifier.submissionpath | gsbs_sp/838 | |
dc.contributor.department | Department of Physiology | |
dc.source.pages | 11-24 | |
dc.contributor.student | James McNally |