Dynamic binding mode of a Synaptotagmin-1-SNARE complex in solution

Kyle D Brewer; Taulant Bacaj; Andrea Cavalli; Carlo Camilloni; James D Swarbrick; Jin Liu; Amy Zhou; Peng Zhou; Nicholas Barlow; Junjie Xu; Alpay B Seven; Eric A Prinslow; Rashmi Voleti; Daniel Häussinger; Alexandre M J J Bonvin; Diana R Tomchick; Michele Vendruscolo; Bim Graham; Thomas C Südhof; Josep Rizo

doi:10.1038/nsmb.3035

Dynamic binding mode of a Synaptotagmin-1-SNARE complex in solution

Kyle D Brewer
, Taulant Bacaj
, Andrea Cavalli
, Carlo Camilloni
, James D Swarbrick
, Jin Liu
, Amy Zhou
, Peng Zhou
, Nicholas Barlow
, Junjie Xu
, Alpay B Seven
, Eric A Prinslow
, Rashmi Voleti
, Daniel Häussinger
, Alexandre M J J Bonvin
, Diana R Tomchick
, Michele Vendruscolo
, Bim Graham
, Thomas C Südhof
, Josep Rizo

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

Rapid neurotransmitter release depends on the Ca2+ sensor Synaptotagmin-1 (Syt1) and the SNARE complex formed by synaptobrevin, syntaxin-1 and SNAP-25. How Syt1 triggers release has been unclear, partly because elucidating high-resolution structures of Syt1-SNARE complexes has been challenging. An NMR approach based on lanthanide-induced pseudocontact shifts now reveals a dynamic binding mode in which basic residues in the concave side of the Syt1 C2B-domain β-sandwich interact with a polyacidic region of the SNARE complex formed by syntaxin-1 and SNAP-25. The physiological relevance of this dynamic structural model is supported by mutations in basic residues of Syt1 that markedly impair SNARE-complex binding in vitro and Syt1 function in neurons. Mutations with milder effects on binding have correspondingly milder effects on Syt1 function. Our results support a model whereby dynamic interaction facilitates cooperation between Syt1 and the SNAREs in inducing membrane fusion.

Original language	English
Pages (from-to)	555-64
Number of pages	10
Journal	Nature Structural and Molecular Biology
Volume	22
Issue number	7
DOIs	https://doi.org/10.1038/nsmb.3035
Publication status	Published - Jul 2015

Keywords

Animals
Cells, Cultured
Humans
Mice, Inbred C57BL
Models, Molecular
Neurons
Nuclear Magnetic Resonance, Biomolecular
Protein Binding
Protein Structure, Tertiary
Rats
SNARE Proteins
Synaptotagmin I

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Brewer, K. D., Bacaj, T., Cavalli, A., Camilloni, C., Swarbrick, J. D., Liu, J., Zhou, A., Zhou, P., Barlow, N., Xu, J., Seven, A. B., Prinslow, E. A., Voleti, R., Häussinger, D., Bonvin, A. M. J. J., Tomchick, D. R., Vendruscolo, M., Graham, B., Südhof, T. C., & Rizo, J. (2015). Dynamic binding mode of a Synaptotagmin-1-SNARE complex in solution. Nature Structural and Molecular Biology, 22(7), 555-64. https://doi.org/10.1038/nsmb.3035

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title = "Dynamic binding mode of a Synaptotagmin-1-SNARE complex in solution",

abstract = "Rapid neurotransmitter release depends on the Ca2+ sensor Synaptotagmin-1 (Syt1) and the SNARE complex formed by synaptobrevin, syntaxin-1 and SNAP-25. How Syt1 triggers release has been unclear, partly because elucidating high-resolution structures of Syt1-SNARE complexes has been challenging. An NMR approach based on lanthanide-induced pseudocontact shifts now reveals a dynamic binding mode in which basic residues in the concave side of the Syt1 C2B-domain β-sandwich interact with a polyacidic region of the SNARE complex formed by syntaxin-1 and SNAP-25. The physiological relevance of this dynamic structural model is supported by mutations in basic residues of Syt1 that markedly impair SNARE-complex binding in vitro and Syt1 function in neurons. Mutations with milder effects on binding have correspondingly milder effects on Syt1 function. Our results support a model whereby dynamic interaction facilitates cooperation between Syt1 and the SNAREs in inducing membrane fusion.",

keywords = "Animals, Cells, Cultured, Humans, Mice, Inbred C57BL, Models, Molecular, Neurons, Nuclear Magnetic Resonance, Biomolecular, Protein Binding, Protein Structure, Tertiary, Rats, SNARE Proteins, Synaptotagmin I",

author = "Brewer, \{Kyle D\} and Taulant Bacaj and Andrea Cavalli and Carlo Camilloni and Swarbrick, \{James D\} and Jin Liu and Amy Zhou and Peng Zhou and Nicholas Barlow and Junjie Xu and Seven, \{Alpay B\} and Prinslow, \{Eric A\} and Rashmi Voleti and Daniel H{\"a}ussinger and Bonvin, \{Alexandre M J J\} and Tomchick, \{Diana R\} and Michele Vendruscolo and Bim Graham and S{\"u}dhof, \{Thomas C\} and Josep Rizo",

year = "2015",

month = jul,

doi = "10.1038/nsmb.3035",

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pages = "555--64",

journal = "Nature Structural and Molecular Biology",

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Brewer, KD, Bacaj, T, Cavalli, A, Camilloni, C, Swarbrick, JD, Liu, J, Zhou, A, Zhou, P, Barlow, N, Xu, J, Seven, AB, Prinslow, EA, Voleti, R, Häussinger, D, Bonvin, AMJJ, Tomchick, DR, Vendruscolo, M, Graham, B, Südhof, TC & Rizo, J 2015, 'Dynamic binding mode of a Synaptotagmin-1-SNARE complex in solution', Nature Structural and Molecular Biology, vol. 22, no. 7, pp. 555-64. https://doi.org/10.1038/nsmb.3035

TY - JOUR

T1 - Dynamic binding mode of a Synaptotagmin-1-SNARE complex in solution

AU - Brewer, Kyle D

AU - Bacaj, Taulant

AU - Cavalli, Andrea

AU - Camilloni, Carlo

AU - Swarbrick, James D

AU - Liu, Jin

AU - Zhou, Amy

AU - Zhou, Peng

AU - Barlow, Nicholas

AU - Xu, Junjie

AU - Seven, Alpay B

AU - Prinslow, Eric A

AU - Voleti, Rashmi

AU - Häussinger, Daniel

AU - Bonvin, Alexandre M J J

AU - Tomchick, Diana R

AU - Vendruscolo, Michele

AU - Graham, Bim

AU - Südhof, Thomas C

AU - Rizo, Josep

PY - 2015/7

Y1 - 2015/7

N2 - Rapid neurotransmitter release depends on the Ca2+ sensor Synaptotagmin-1 (Syt1) and the SNARE complex formed by synaptobrevin, syntaxin-1 and SNAP-25. How Syt1 triggers release has been unclear, partly because elucidating high-resolution structures of Syt1-SNARE complexes has been challenging. An NMR approach based on lanthanide-induced pseudocontact shifts now reveals a dynamic binding mode in which basic residues in the concave side of the Syt1 C2B-domain β-sandwich interact with a polyacidic region of the SNARE complex formed by syntaxin-1 and SNAP-25. The physiological relevance of this dynamic structural model is supported by mutations in basic residues of Syt1 that markedly impair SNARE-complex binding in vitro and Syt1 function in neurons. Mutations with milder effects on binding have correspondingly milder effects on Syt1 function. Our results support a model whereby dynamic interaction facilitates cooperation between Syt1 and the SNAREs in inducing membrane fusion.

AB - Rapid neurotransmitter release depends on the Ca2+ sensor Synaptotagmin-1 (Syt1) and the SNARE complex formed by synaptobrevin, syntaxin-1 and SNAP-25. How Syt1 triggers release has been unclear, partly because elucidating high-resolution structures of Syt1-SNARE complexes has been challenging. An NMR approach based on lanthanide-induced pseudocontact shifts now reveals a dynamic binding mode in which basic residues in the concave side of the Syt1 C2B-domain β-sandwich interact with a polyacidic region of the SNARE complex formed by syntaxin-1 and SNAP-25. The physiological relevance of this dynamic structural model is supported by mutations in basic residues of Syt1 that markedly impair SNARE-complex binding in vitro and Syt1 function in neurons. Mutations with milder effects on binding have correspondingly milder effects on Syt1 function. Our results support a model whereby dynamic interaction facilitates cooperation between Syt1 and the SNAREs in inducing membrane fusion.

KW - Animals

KW - Cells, Cultured

KW - Humans

KW - Mice, Inbred C57BL

KW - Models, Molecular

KW - Neurons

KW - Nuclear Magnetic Resonance, Biomolecular

KW - Protein Binding

KW - Protein Structure, Tertiary

KW - Rats

KW - SNARE Proteins

KW - Synaptotagmin I

U2 - 10.1038/nsmb.3035

DO - 10.1038/nsmb.3035

M3 - Article

C2 - 26030874

SN - 1545-9993

VL - 22

SP - 555

EP - 564

JO - Nature Structural and Molecular Biology

JF - Nature Structural and Molecular Biology

IS - 7

ER -

Dynamic binding mode of a Synaptotagmin-1-SNARE complex in solution

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Keywords

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