A Coiled Coil Trigger Site Is Essential for Rapid Binding of Synaptobrevin to the SNARE Acceptor Complex
Exocytosis from synaptic vesicles is driven by stepwise formation of a tight α-helical complex between the fusing membranes. The complex is composed of the three SNAREs: synaptobrevin 2, SNAP-25, and syntaxin 1a. An important step in complex formation is fast binding of vesicular synaptobrevin to th...
Gespeichert in:
| Veröffentlicht in: | The Journal of biological chemistry 2010-07, Vol.285 (28), p.21549-21559 |
|---|---|
| Hauptverfasser: | , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 21559 |
|---|---|
| container_issue | 28 |
| container_start_page | 21549 |
| container_title | The Journal of biological chemistry |
| container_volume | 285 |
| creator | Wiederhold, Katrin Kloepper, Tobias H. Walter, Alexander M. Stein, Alexander Kienle, Nickias Sørensen, Jakob B. Fasshauer, Dirk |
| description | Exocytosis from synaptic vesicles is driven by stepwise formation of a tight α-helical complex between the fusing membranes. The complex is composed of the three SNAREs: synaptobrevin 2, SNAP-25, and syntaxin 1a. An important step in complex formation is fast binding of vesicular synaptobrevin to the preformed syntaxin 1·SNAP-25 dimer. Exactly how this step relates to neurotransmitter release is not well understood. Here, we combined different approaches to gain insights into this reaction. Using computational methods, we identified a stretch in synaptobrevin 2 that may function as a coiled coil “trigger site.” This site is also present in many synaptobrevin homologs functioning in other trafficking steps. Point mutations in this stretch inhibited binding to the syntaxin 1·SNAP-25 dimer and slowed fusion of liposomes. Moreover, the point mutations severely inhibited secretion from chromaffin cells. Altogether, this demonstrates that the trigger site in synaptobrevin is crucial for productive SNARE zippering. |
| doi_str_mv | 10.1074/jbc.M110.105148 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_2898431</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S002192582060182X</els_id><sourcerecordid>754537684</sourcerecordid><originalsourceid>FETCH-LOGICAL-c498t-58fcd302899837a4716f11a0cb924e606dd7fd810d8be9fd204e4fea75fb58483</originalsourceid><addsrcrecordid>eNqFkU1v1DAQhi0EokvhzA1845TWju3EuSAtqwUqlSJ1W4mb5djjrKtsHOzsiv573KZUcED1ZWTNM-98vAi9peSEkpqf3rTm5Bu9_wnK5TO0oESyggn64zlaEFLSoimFPEKvUroh-fGGvkRHJeGkkiVdoO0Sr4Lvwd4HfBV910HEGz8BPkt4nRIMk9c9diHiSz16iz_5wfqhw8Hhze2gxym0EQ5-wFPA0xbw5mJ5ucZLYyCnYtbdjT38eo1eON0nePMQj9H15_XV6mtx_v3L2Wp5XhjeyKkQ0hnLSCmbRrJa85pWjlJNTNuUHCpSWVs7KymxsoXG2bwJcAe6Fq4Vkkt2jD7OuuO-3YE1efyoezVGv9PxVgXt1b-ZwW9VFw4qt5Sc0Szw4UEghp97SJPa-WSg7_UAYZ9ULbhgdZXZJ0nGKiZEJTJ5OpMmhpQiuMd5KFF3RqpspLozUs1G5op3f6_xyP9xLgPvZ8DpoHQXfVLXm5JQRqisaC2aTDQzAfncBw9RJeNhMGB9BDMpG_x_2_8Gsz-13g</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>733635565</pqid></control><display><type>article</type><title>A Coiled Coil Trigger Site Is Essential for Rapid Binding of Synaptobrevin to the SNARE Acceptor Complex</title><source>MEDLINE</source><source>EZB-FREE-00999 freely available EZB journals</source><source>PubMed Central</source><source>Alma/SFX Local Collection</source><creator>Wiederhold, Katrin ; Kloepper, Tobias H. ; Walter, Alexander M. ; Stein, Alexander ; Kienle, Nickias ; Sørensen, Jakob B. ; Fasshauer, Dirk</creator><creatorcontrib>Wiederhold, Katrin ; Kloepper, Tobias H. ; Walter, Alexander M. ; Stein, Alexander ; Kienle, Nickias ; Sørensen, Jakob B. ; Fasshauer, Dirk</creatorcontrib><description>Exocytosis from synaptic vesicles is driven by stepwise formation of a tight α-helical complex between the fusing membranes. The complex is composed of the three SNAREs: synaptobrevin 2, SNAP-25, and syntaxin 1a. An important step in complex formation is fast binding of vesicular synaptobrevin to the preformed syntaxin 1·SNAP-25 dimer. Exactly how this step relates to neurotransmitter release is not well understood. Here, we combined different approaches to gain insights into this reaction. Using computational methods, we identified a stretch in synaptobrevin 2 that may function as a coiled coil “trigger site.” This site is also present in many synaptobrevin homologs functioning in other trafficking steps. Point mutations in this stretch inhibited binding to the syntaxin 1·SNAP-25 dimer and slowed fusion of liposomes. Moreover, the point mutations severely inhibited secretion from chromaffin cells. Altogether, this demonstrates that the trigger site in synaptobrevin is crucial for productive SNARE zippering.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1074/jbc.M110.105148</identifier><identifier>PMID: 20406821</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Amino Acid Motifs ; Animals ; Binding Sites ; Calcium - chemistry ; Calorimetry - methods ; Chromaffin Cells - metabolism ; Coiled Coil ; Dimerization ; Electrophysiology - methods ; Intracellular Trafficking ; Isothermal Titration Calorimetry ; Liposomes - chemistry ; Membrane Biology ; Membrane Fusion ; Mice ; Neurobiology ; Neurotransmitter Agents - metabolism ; Neurotransmitters ; Point Mutation ; Protein Structure, Tertiary ; R-SNARE Proteins - chemistry ; Rats ; SNARE Protein ; SNARE Proteins - chemistry ; Synapses ; Vesicles</subject><ispartof>The Journal of biological chemistry, 2010-07, Vol.285 (28), p.21549-21559</ispartof><rights>2010 © 2010 ASBMB. Currently published by Elsevier Inc; originally published by American Society for Biochemistry and Molecular Biology.</rights><rights>2010 by The American Society for Biochemistry and Molecular Biology, Inc.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c498t-58fcd302899837a4716f11a0cb924e606dd7fd810d8be9fd204e4fea75fb58483</citedby><cites>FETCH-LOGICAL-c498t-58fcd302899837a4716f11a0cb924e606dd7fd810d8be9fd204e4fea75fb58483</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2898431/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2898431/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,27915,27916,53782,53784</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20406821$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Wiederhold, Katrin</creatorcontrib><creatorcontrib>Kloepper, Tobias H.</creatorcontrib><creatorcontrib>Walter, Alexander M.</creatorcontrib><creatorcontrib>Stein, Alexander</creatorcontrib><creatorcontrib>Kienle, Nickias</creatorcontrib><creatorcontrib>Sørensen, Jakob B.</creatorcontrib><creatorcontrib>Fasshauer, Dirk</creatorcontrib><title>A Coiled Coil Trigger Site Is Essential for Rapid Binding of Synaptobrevin to the SNARE Acceptor Complex</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>Exocytosis from synaptic vesicles is driven by stepwise formation of a tight α-helical complex between the fusing membranes. The complex is composed of the three SNAREs: synaptobrevin 2, SNAP-25, and syntaxin 1a. An important step in complex formation is fast binding of vesicular synaptobrevin to the preformed syntaxin 1·SNAP-25 dimer. Exactly how this step relates to neurotransmitter release is not well understood. Here, we combined different approaches to gain insights into this reaction. Using computational methods, we identified a stretch in synaptobrevin 2 that may function as a coiled coil “trigger site.” This site is also present in many synaptobrevin homologs functioning in other trafficking steps. Point mutations in this stretch inhibited binding to the syntaxin 1·SNAP-25 dimer and slowed fusion of liposomes. Moreover, the point mutations severely inhibited secretion from chromaffin cells. Altogether, this demonstrates that the trigger site in synaptobrevin is crucial for productive SNARE zippering.</description><subject>Amino Acid Motifs</subject><subject>Animals</subject><subject>Binding Sites</subject><subject>Calcium - chemistry</subject><subject>Calorimetry - methods</subject><subject>Chromaffin Cells - metabolism</subject><subject>Coiled Coil</subject><subject>Dimerization</subject><subject>Electrophysiology - methods</subject><subject>Intracellular Trafficking</subject><subject>Isothermal Titration Calorimetry</subject><subject>Liposomes - chemistry</subject><subject>Membrane Biology</subject><subject>Membrane Fusion</subject><subject>Mice</subject><subject>Neurobiology</subject><subject>Neurotransmitter Agents - metabolism</subject><subject>Neurotransmitters</subject><subject>Point Mutation</subject><subject>Protein Structure, Tertiary</subject><subject>R-SNARE Proteins - chemistry</subject><subject>Rats</subject><subject>SNARE Protein</subject><subject>SNARE Proteins - chemistry</subject><subject>Synapses</subject><subject>Vesicles</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkU1v1DAQhi0EokvhzA1845TWju3EuSAtqwUqlSJ1W4mb5djjrKtsHOzsiv573KZUcED1ZWTNM-98vAi9peSEkpqf3rTm5Bu9_wnK5TO0oESyggn64zlaEFLSoimFPEKvUroh-fGGvkRHJeGkkiVdoO0Sr4Lvwd4HfBV910HEGz8BPkt4nRIMk9c9diHiSz16iz_5wfqhw8Hhze2gxym0EQ5-wFPA0xbw5mJ5ucZLYyCnYtbdjT38eo1eON0nePMQj9H15_XV6mtx_v3L2Wp5XhjeyKkQ0hnLSCmbRrJa85pWjlJNTNuUHCpSWVs7KymxsoXG2bwJcAe6Fq4Vkkt2jD7OuuO-3YE1efyoezVGv9PxVgXt1b-ZwW9VFw4qt5Sc0Szw4UEghp97SJPa-WSg7_UAYZ9ULbhgdZXZJ0nGKiZEJTJ5OpMmhpQiuMd5KFF3RqpspLozUs1G5op3f6_xyP9xLgPvZ8DpoHQXfVLXm5JQRqisaC2aTDQzAfncBw9RJeNhMGB9BDMpG_x_2_8Gsz-13g</recordid><startdate>20100709</startdate><enddate>20100709</enddate><creator>Wiederhold, Katrin</creator><creator>Kloepper, Tobias H.</creator><creator>Walter, Alexander M.</creator><creator>Stein, Alexander</creator><creator>Kienle, Nickias</creator><creator>Sørensen, Jakob B.</creator><creator>Fasshauer, Dirk</creator><general>Elsevier Inc</general><general>American Society for Biochemistry and Molecular Biology</general><scope>6I.</scope><scope>AAFTH</scope><scope>FBQ</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>7TK</scope><scope>5PM</scope></search><sort><creationdate>20100709</creationdate><title>A Coiled Coil Trigger Site Is Essential for Rapid Binding of Synaptobrevin to the SNARE Acceptor Complex</title><author>Wiederhold, Katrin ; Kloepper, Tobias H. ; Walter, Alexander M. ; Stein, Alexander ; Kienle, Nickias ; Sørensen, Jakob B. ; Fasshauer, Dirk</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c498t-58fcd302899837a4716f11a0cb924e606dd7fd810d8be9fd204e4fea75fb58483</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>Amino Acid Motifs</topic><topic>Animals</topic><topic>Binding Sites</topic><topic>Calcium - chemistry</topic><topic>Calorimetry - methods</topic><topic>Chromaffin Cells - metabolism</topic><topic>Coiled Coil</topic><topic>Dimerization</topic><topic>Electrophysiology - methods</topic><topic>Intracellular Trafficking</topic><topic>Isothermal Titration Calorimetry</topic><topic>Liposomes - chemistry</topic><topic>Membrane Biology</topic><topic>Membrane Fusion</topic><topic>Mice</topic><topic>Neurobiology</topic><topic>Neurotransmitter Agents - metabolism</topic><topic>Neurotransmitters</topic><topic>Point Mutation</topic><topic>Protein Structure, Tertiary</topic><topic>R-SNARE Proteins - chemistry</topic><topic>Rats</topic><topic>SNARE Protein</topic><topic>SNARE Proteins - chemistry</topic><topic>Synapses</topic><topic>Vesicles</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wiederhold, Katrin</creatorcontrib><creatorcontrib>Kloepper, Tobias H.</creatorcontrib><creatorcontrib>Walter, Alexander M.</creatorcontrib><creatorcontrib>Stein, Alexander</creatorcontrib><creatorcontrib>Kienle, Nickias</creatorcontrib><creatorcontrib>Sørensen, Jakob B.</creatorcontrib><creatorcontrib>Fasshauer, Dirk</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Neurosciences Abstracts</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wiederhold, Katrin</au><au>Kloepper, Tobias H.</au><au>Walter, Alexander M.</au><au>Stein, Alexander</au><au>Kienle, Nickias</au><au>Sørensen, Jakob B.</au><au>Fasshauer, Dirk</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A Coiled Coil Trigger Site Is Essential for Rapid Binding of Synaptobrevin to the SNARE Acceptor Complex</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>2010-07-09</date><risdate>2010</risdate><volume>285</volume><issue>28</issue><spage>21549</spage><epage>21559</epage><pages>21549-21559</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><abstract>Exocytosis from synaptic vesicles is driven by stepwise formation of a tight α-helical complex between the fusing membranes. The complex is composed of the three SNAREs: synaptobrevin 2, SNAP-25, and syntaxin 1a. An important step in complex formation is fast binding of vesicular synaptobrevin to the preformed syntaxin 1·SNAP-25 dimer. Exactly how this step relates to neurotransmitter release is not well understood. Here, we combined different approaches to gain insights into this reaction. Using computational methods, we identified a stretch in synaptobrevin 2 that may function as a coiled coil “trigger site.” This site is also present in many synaptobrevin homologs functioning in other trafficking steps. Point mutations in this stretch inhibited binding to the syntaxin 1·SNAP-25 dimer and slowed fusion of liposomes. Moreover, the point mutations severely inhibited secretion from chromaffin cells. Altogether, this demonstrates that the trigger site in synaptobrevin is crucial for productive SNARE zippering.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>20406821</pmid><doi>10.1074/jbc.M110.105148</doi><tpages>11</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0021-9258 |
| ispartof | The Journal of biological chemistry, 2010-07, Vol.285 (28), p.21549-21559 |
| issn | 0021-9258 1083-351X |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_2898431 |
| source | MEDLINE; EZB-FREE-00999 freely available EZB journals; PubMed Central; Alma/SFX Local Collection |
| subjects | Amino Acid Motifs Animals Binding Sites Calcium - chemistry Calorimetry - methods Chromaffin Cells - metabolism Coiled Coil Dimerization Electrophysiology - methods Intracellular Trafficking Isothermal Titration Calorimetry Liposomes - chemistry Membrane Biology Membrane Fusion Mice Neurobiology Neurotransmitter Agents - metabolism Neurotransmitters Point Mutation Protein Structure, Tertiary R-SNARE Proteins - chemistry Rats SNARE Protein SNARE Proteins - chemistry Synapses Vesicles |
| title | A Coiled Coil Trigger Site Is Essential for Rapid Binding of Synaptobrevin to the SNARE Acceptor Complex |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-15T00%3A52%3A29IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=A%20Coiled%20Coil%20Trigger%20Site%20Is%20Essential%20for%20Rapid%20Binding%20of%20Synaptobrevin%20to%20the%20SNARE%20Acceptor%20Complex&rft.jtitle=The%20Journal%20of%20biological%20chemistry&rft.au=Wiederhold,%20Katrin&rft.date=2010-07-09&rft.volume=285&rft.issue=28&rft.spage=21549&rft.epage=21559&rft.pages=21549-21559&rft.issn=0021-9258&rft.eissn=1083-351X&rft_id=info:doi/10.1074/jbc.M110.105148&rft_dat=%3Cproquest_pubme%3E754537684%3C/proquest_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=733635565&rft_id=info:pmid/20406821&rft_els_id=S002192582060182X&rfr_iscdi=true |