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PMID: 26030874 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

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

Nature structural & molecular biology ·Vol. 22 ·No. 7 ·2015-07-00 ·Pages 555-64

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 AM, Tomchick DR, Vendruscolo M, Graham B, Südhof TC, Rizo J

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.

MeSH Terms
Animals Cells, Cultured Humans Mice, Inbred C57BL Models, Molecular Neurons/metabolism Nuclear Magnetic Resonance, Biomolecular Protein Binding Protein Structure, Tertiary Rats SNARE Proteins/chemistry,metabolism Synaptotagmin I/chemistry,metabolism
Chemicals
SNARE Proteins Synaptotagmin I
Authors & Affiliations
20 authors, click to expand affiliations / ORCID
Brewer Kyle D
1] Department of Biophysics, University of Texas Southwestern Medical Center, Dallas, Texas, USA. [2] Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, Texas, USA. [3] Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
Bacaj Taulant
1] Department of Molecular and Cellular Physiology, Stanford University Medical School, Stanford, California, USA. [2] Howard Hughes Medical Institute, Stanford University Medical School, Stanford, California, USA.
Cavalli Andrea
1] Institute for Research in Biomedicine, Bellinzona, Switzerland. [2] Department of Chemistry, University of Cambridge, Cambridge, UK.
Camilloni Carlo ORCID
Department of Chemistry, University of Cambridge, Cambridge, UK.
Swarbrick James D
Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria, Australia.
Liu Jin
1] Department of Biophysics, University of Texas Southwestern Medical Center, Dallas, Texas, USA. [2] Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, Texas, USA. [3] Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, Texas, USA. [4] Department of Chemistry, Southern Methodist University, Dallas, Texas, USA.
Zhou Amy
1] Department of Biophysics, University of Texas Southwestern Medical Center, Dallas, Texas, USA. [2] Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, Texas, USA. [3] Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
Zhou Peng
1] Department of Molecular and Cellular Physiology, Stanford University Medical School, Stanford, California, USA. [2] Howard Hughes Medical Institute, Stanford University Medical School, Stanford, California, USA.
Barlow Nicholas
Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria, Australia.
Xu Junjie
1] Department of Biophysics, University of Texas Southwestern Medical Center, Dallas, Texas, USA. [2] Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, Texas, USA. [3] Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
Seven Alpay B
1] Department of Biophysics, University of Texas Southwestern Medical Center, Dallas, Texas, USA. [2] Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, Texas, USA. [3] Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
Prinslow Eric A
1] Department of Biophysics, University of Texas Southwestern Medical Center, Dallas, Texas, USA. [2] Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, Texas, USA. [3] Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
Voleti Rashmi
1] Department of Biophysics, University of Texas Southwestern Medical Center, Dallas, Texas, USA. [2] Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, Texas, USA. [3] Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
Häussinger Daniel
Department of Chemistry, University of Basel, Basel, Switzerland.
Bonvin Alexandre M J J
Bijvoet Center for Biomolecular Research, Faculty of Science, Utrecht University, Utrecht, the Netherlands.
Tomchick Diana R
1] Department of Biophysics, University of Texas Southwestern Medical Center, Dallas, Texas, USA. [2] Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
Vendruscolo Michele
Department of Chemistry, University of Cambridge, Cambridge, UK.
Graham Bim
Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria, Australia.
Südhof Thomas C
1] Department of Molecular and Cellular Physiology, Stanford University Medical School, Stanford, California, USA. [2] Howard Hughes Medical Institute, Stanford University Medical School, Stanford, California, USA.
Rizo Josep
1] Department of Biophysics, University of Texas Southwestern Medical Center, Dallas, Texas, USA. [2] Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, Texas, USA. [3] Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
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Article Info
Journal
Nature structural & molecular biology
Abbr.
Nat Struct Mol Biol
ISSN
1545-9985
DOI
10.1038/nsmb.3035
Published
2015-07-00
Epub
2015-00-01
Pages
555-64
Language
English
Region
United States
NLM ID
101186374
PMCID
PMC4496268
Subset
IM
Grants
NINDS NIH HHS · K99NS087086 · United States
NINDS NIH HHS · K99 NS087086 · United States
NINDS NIH HHS · R01 NS040944 · United States
NIGMS NIH HHS · T32 GM008297 · United States
NINDS NIH HHS · NS040944 · United States
NCRR NIH HHS · S10 RR026461 · United States
Corrections
CommentIn
PMID: 26150331
External Links
PubMed source DOI: 10.1038/nsmb.3035 PMC Full Text View journal details
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