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Regulation of syntaxin1Aâmunc18 complex for SNARE pairing in HEK293 cells
The formation and dissolution of SNARE protein complexes is essential for Ca 2+ -triggered fusion of neurotransmitter-filled vesicles at the presynaptic membrane. Among the pre-synaptic SNARE proteins, the activation of the Q-SNARE syntaxin1A is a critical event for SNARE complex formation. Activati...
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Published in: | The Journal of physiology 2004-08, Vol.558 (3), p.857 |
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container_title | The Journal of physiology |
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creator | Svetlana E. Gladycheva Chi S. Ho Yue Ying F. Lee Edward L. Stuenkel |
description | The formation and dissolution of SNARE protein complexes is essential for Ca 2+ -triggered fusion of neurotransmitter-filled vesicles at the presynaptic membrane. Among the pre-synaptic SNARE proteins,
the activation of the Q-SNARE syntaxin1A is a critical event for SNARE complex formation. Activation requires syntaxin1A to
transit from a munc18-bound non-interacting state to one competent for SNARE binding. The molecular mechanisms that regulate
this transition remain unclear. The propensity of syntaxin1A to promote voltage-dependent steady-state inactivation of N-type
Ca 2+ channels and accelerate their entry into inactivation was used in a heterologous cell expression system to elucidate regulation
of syntaxin1A proteinâprotein interactions. We report that coexpression of munc18 eliminated the promoting effect of syntaxin1A
on inactivation. This effect of munc18 was completely disrupted by coexpression of munc13-1, but not munc13-2 or munc13-3.
Also, since expression of munc13-1 with syntaxin1A resulted in an inactivation phenotype identical to that of munc18 with
syntaxin1A, the action of munc13-1 on the munc18âsyntaxin1A complex was functionally unique and did not result from competitive
binding interactions. Furthermore, munc13 expressed with syntaxin1A and munc18 promoted redistribution of a cytosolic SNAP25
mutant to the membrane, a result indicative of syntaxin1AâSNAP25 SNARE pairing. These data demonstrate an important role of
munc13 to control the proteinâprotein interactions of syntaxin1A in vivo, and support munc13 as critical to dissociating syntaxin1Aâmunc18 complexes and making syntaxin1A available for SNARE interactions. |
doi_str_mv | 10.1113/jphysiol.2004.067249 |
format | article |
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the activation of the Q-SNARE syntaxin1A is a critical event for SNARE complex formation. Activation requires syntaxin1A to
transit from a munc18-bound non-interacting state to one competent for SNARE binding. The molecular mechanisms that regulate
this transition remain unclear. The propensity of syntaxin1A to promote voltage-dependent steady-state inactivation of N-type
Ca 2+ channels and accelerate their entry into inactivation was used in a heterologous cell expression system to elucidate regulation
of syntaxin1A proteinâprotein interactions. We report that coexpression of munc18 eliminated the promoting effect of syntaxin1A
on inactivation. This effect of munc18 was completely disrupted by coexpression of munc13-1, but not munc13-2 or munc13-3.
Also, since expression of munc13-1 with syntaxin1A resulted in an inactivation phenotype identical to that of munc18 with
syntaxin1A, the action of munc13-1 on the munc18âsyntaxin1A complex was functionally unique and did not result from competitive
binding interactions. Furthermore, munc13 expressed with syntaxin1A and munc18 promoted redistribution of a cytosolic SNAP25
mutant to the membrane, a result indicative of syntaxin1AâSNAP25 SNARE pairing. These data demonstrate an important role of
munc13 to control the proteinâprotein interactions of syntaxin1A in vivo, and support munc13 as critical to dissociating syntaxin1Aâmunc18 complexes and making syntaxin1A available for SNARE interactions.</description><identifier>ISSN: 0022-3751</identifier><identifier>EISSN: 1469-7793</identifier><identifier>DOI: 10.1113/jphysiol.2004.067249</identifier><identifier>PMID: 15218059</identifier><language>eng</language><publisher>The Physiological Society</publisher><ispartof>The Journal of physiology, 2004-08, Vol.558 (3), p.857</ispartof><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids></links><search><creatorcontrib>Svetlana E. Gladycheva</creatorcontrib><creatorcontrib>Chi S. Ho</creatorcontrib><creatorcontrib>Yue Ying F. Lee</creatorcontrib><creatorcontrib>Edward L. Stuenkel</creatorcontrib><title>Regulation of syntaxin1Aâmunc18 complex for SNARE pairing in HEK293 cells</title><title>The Journal of physiology</title><description>The formation and dissolution of SNARE protein complexes is essential for Ca 2+ -triggered fusion of neurotransmitter-filled vesicles at the presynaptic membrane. Among the pre-synaptic SNARE proteins,
the activation of the Q-SNARE syntaxin1A is a critical event for SNARE complex formation. Activation requires syntaxin1A to
transit from a munc18-bound non-interacting state to one competent for SNARE binding. The molecular mechanisms that regulate
this transition remain unclear. The propensity of syntaxin1A to promote voltage-dependent steady-state inactivation of N-type
Ca 2+ channels and accelerate their entry into inactivation was used in a heterologous cell expression system to elucidate regulation
of syntaxin1A proteinâprotein interactions. We report that coexpression of munc18 eliminated the promoting effect of syntaxin1A
on inactivation. This effect of munc18 was completely disrupted by coexpression of munc13-1, but not munc13-2 or munc13-3.
Also, since expression of munc13-1 with syntaxin1A resulted in an inactivation phenotype identical to that of munc18 with
syntaxin1A, the action of munc13-1 on the munc18âsyntaxin1A complex was functionally unique and did not result from competitive
binding interactions. Furthermore, munc13 expressed with syntaxin1A and munc18 promoted redistribution of a cytosolic SNAP25
mutant to the membrane, a result indicative of syntaxin1AâSNAP25 SNARE pairing. These data demonstrate an important role of
munc13 to control the proteinâprotein interactions of syntaxin1A in vivo, and support munc13 as critical to dissociating syntaxin1Aâmunc18 complexes and making syntaxin1A available for SNARE interactions.</description><issn>0022-3751</issn><issn>1469-7793</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2004</creationdate><recordtype>article</recordtype><sourceid/><recordid>eNqNzj9OwzAUgHELgWj4cwOGtzElvGfHdTxWKKgSUofCHkWRk7hy7ShORbNxB27AUcrFGMoBmL7lN3yMPRBmRCSedkM_RxtcxhHzDJeK5_qCJZQvdaqUFpcsQeQ8FUrSgt3EuEMkgVpfswVJTgVKnbDN1nQHV082eAgtxNlP9dF6Wv18nz5PX_uDb6iAJuwHZ47QhhHeNqttCUNtR-s7sB7W5SvXAhrjXLxjV23torn_6y17fCnfn9dpb7v-w46mOl_H0FgzzZWURSWqQirxf_kLBvJNDA</recordid><startdate>20040801</startdate><enddate>20040801</enddate><creator>Svetlana E. Gladycheva</creator><creator>Chi S. Ho</creator><creator>Yue Ying F. Lee</creator><creator>Edward L. Stuenkel</creator><general>The Physiological Society</general><scope/></search><sort><creationdate>20040801</creationdate><title>Regulation of syntaxin1Aâmunc18 complex for SNARE pairing in HEK293 cells</title><author>Svetlana E. Gladycheva ; Chi S. Ho ; Yue Ying F. Lee ; Edward L. Stuenkel</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-highwire_physiosociety_558_3_8573</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2004</creationdate><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Svetlana E. Gladycheva</creatorcontrib><creatorcontrib>Chi S. Ho</creatorcontrib><creatorcontrib>Yue Ying F. Lee</creatorcontrib><creatorcontrib>Edward L. Stuenkel</creatorcontrib><jtitle>The Journal of physiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Svetlana E. Gladycheva</au><au>Chi S. Ho</au><au>Yue Ying F. Lee</au><au>Edward L. Stuenkel</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Regulation of syntaxin1Aâmunc18 complex for SNARE pairing in HEK293 cells</atitle><jtitle>The Journal of physiology</jtitle><date>2004-08-01</date><risdate>2004</risdate><volume>558</volume><issue>3</issue><spage>857</spage><pages>857-</pages><issn>0022-3751</issn><eissn>1469-7793</eissn><abstract>The formation and dissolution of SNARE protein complexes is essential for Ca 2+ -triggered fusion of neurotransmitter-filled vesicles at the presynaptic membrane. Among the pre-synaptic SNARE proteins,
the activation of the Q-SNARE syntaxin1A is a critical event for SNARE complex formation. Activation requires syntaxin1A to
transit from a munc18-bound non-interacting state to one competent for SNARE binding. The molecular mechanisms that regulate
this transition remain unclear. The propensity of syntaxin1A to promote voltage-dependent steady-state inactivation of N-type
Ca 2+ channels and accelerate their entry into inactivation was used in a heterologous cell expression system to elucidate regulation
of syntaxin1A proteinâprotein interactions. We report that coexpression of munc18 eliminated the promoting effect of syntaxin1A
on inactivation. This effect of munc18 was completely disrupted by coexpression of munc13-1, but not munc13-2 or munc13-3.
Also, since expression of munc13-1 with syntaxin1A resulted in an inactivation phenotype identical to that of munc18 with
syntaxin1A, the action of munc13-1 on the munc18âsyntaxin1A complex was functionally unique and did not result from competitive
binding interactions. Furthermore, munc13 expressed with syntaxin1A and munc18 promoted redistribution of a cytosolic SNAP25
mutant to the membrane, a result indicative of syntaxin1AâSNAP25 SNARE pairing. These data demonstrate an important role of
munc13 to control the proteinâprotein interactions of syntaxin1A in vivo, and support munc13 as critical to dissociating syntaxin1Aâmunc18 complexes and making syntaxin1A available for SNARE interactions.</abstract><pub>The Physiological Society</pub><pmid>15218059</pmid><doi>10.1113/jphysiol.2004.067249</doi></addata></record> |
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title | Regulation of syntaxin1Aâmunc18 complex for SNARE pairing in HEK293 cells |
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