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Reconciling isothermal titration calorimetry analyses of interactions between complexin and truncated SNARE complexes

Neurotransmitter release depends on the SNARE complex formed by syntaxin-1, synaptobrevin and SNAP-25, as well as on complexins, which bind to the SNARE complex and play active and inhibitory roles. A crystal structure of a Complexin-I fragment bearing a so-called 'superclamp' mutation bou...

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Autores principales: Prinslow, Eric A, Brautigam, Chad A, Rizo, Josep
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5589412/
https://www.ncbi.nlm.nih.gov/pubmed/28880148
http://dx.doi.org/10.7554/eLife.30286
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author Prinslow, Eric A
Brautigam, Chad A
Rizo, Josep
author_facet Prinslow, Eric A
Brautigam, Chad A
Rizo, Josep
author_sort Prinslow, Eric A
collection PubMed
description Neurotransmitter release depends on the SNARE complex formed by syntaxin-1, synaptobrevin and SNAP-25, as well as on complexins, which bind to the SNARE complex and play active and inhibitory roles. A crystal structure of a Complexin-I fragment bearing a so-called 'superclamp' mutation bound to a truncated SNARE complex lacking the C-terminus of the synaptobrevin SNARE motif (SNAREΔ60) suggested that an 'accessory' α-helix of Complexin-I inhibits release by inserting into the C-terminus of the SNARE complex. Previously, isothermal titration calorimetry (ITC) experiments performed in different laboratories yielded apparently discrepant results in support or against the existence of such binding mode in solution (Trimbuch et al., 2014; Krishnakumar et al., 2015). Here, ITC experiments performed to solve these discrepancies now show that the region containing the Complexin-I accessory helix and preceding N-terminal sequences does interact with SNAREΔ60, but the interaction requires the polybasic juxtamembrane region of syntaxin-1 and is not affected by the superclamp mutation within the experimental error of these experiments.
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spelling pubmed-55894122017-09-11 Reconciling isothermal titration calorimetry analyses of interactions between complexin and truncated SNARE complexes Prinslow, Eric A Brautigam, Chad A Rizo, Josep eLife Structural Biology and Molecular Biophysics Neurotransmitter release depends on the SNARE complex formed by syntaxin-1, synaptobrevin and SNAP-25, as well as on complexins, which bind to the SNARE complex and play active and inhibitory roles. A crystal structure of a Complexin-I fragment bearing a so-called 'superclamp' mutation bound to a truncated SNARE complex lacking the C-terminus of the synaptobrevin SNARE motif (SNAREΔ60) suggested that an 'accessory' α-helix of Complexin-I inhibits release by inserting into the C-terminus of the SNARE complex. Previously, isothermal titration calorimetry (ITC) experiments performed in different laboratories yielded apparently discrepant results in support or against the existence of such binding mode in solution (Trimbuch et al., 2014; Krishnakumar et al., 2015). Here, ITC experiments performed to solve these discrepancies now show that the region containing the Complexin-I accessory helix and preceding N-terminal sequences does interact with SNAREΔ60, but the interaction requires the polybasic juxtamembrane region of syntaxin-1 and is not affected by the superclamp mutation within the experimental error of these experiments. eLife Sciences Publications, Ltd 2017-09-07 /pmc/articles/PMC5589412/ /pubmed/28880148 http://dx.doi.org/10.7554/eLife.30286 Text en © 2017, Prinslow et al http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Structural Biology and Molecular Biophysics
Prinslow, Eric A
Brautigam, Chad A
Rizo, Josep
Reconciling isothermal titration calorimetry analyses of interactions between complexin and truncated SNARE complexes
title Reconciling isothermal titration calorimetry analyses of interactions between complexin and truncated SNARE complexes
title_full Reconciling isothermal titration calorimetry analyses of interactions between complexin and truncated SNARE complexes
title_fullStr Reconciling isothermal titration calorimetry analyses of interactions between complexin and truncated SNARE complexes
title_full_unstemmed Reconciling isothermal titration calorimetry analyses of interactions between complexin and truncated SNARE complexes
title_short Reconciling isothermal titration calorimetry analyses of interactions between complexin and truncated SNARE complexes
title_sort reconciling isothermal titration calorimetry analyses of interactions between complexin and truncated snare complexes
topic Structural Biology and Molecular Biophysics
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5589412/
https://www.ncbi.nlm.nih.gov/pubmed/28880148
http://dx.doi.org/10.7554/eLife.30286
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