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TitleStructural basis for the clamping and Ca activation of SNARE-mediated fusion by synaptotagmin.
Journal, issue, pagesNat Commun, Vol. 10, Issue 1, Page 2413, Year 2019
Publish dateJun 3, 2019
AuthorsKirill Grushin / Jing Wang / Jeff Coleman / James E Rothman / Charles V Sindelar / Shyam S Krishnakumar /
PubMed AbstractSynapotagmin-1 (Syt1) interacts with both SNARE proteins and lipid membranes to synchronize neurotransmitter release to calcium (Ca) influx. Here we report the cryo-electron microscopy structure of ...Synapotagmin-1 (Syt1) interacts with both SNARE proteins and lipid membranes to synchronize neurotransmitter release to calcium (Ca) influx. Here we report the cryo-electron microscopy structure of the Syt1-SNARE complex on anionic-lipid containing membranes. Under resting conditions, the Syt1 C2 domains bind the membrane with a magnesium (Mg)-mediated partial insertion of the aliphatic loops, alongside weak interactions with the anionic lipid headgroups. The C2B domain concurrently interacts the SNARE bundle via the 'primary' interface and is positioned between the SNAREpins and the membrane. In this configuration, Syt1 is projected to sterically delay the complete assembly of the associated SNAREpins and thus, contribute to clamping fusion. This Syt1-SNARE organization is disrupted upon Ca-influx as Syt1 reorients into the membrane, likely displacing the attached SNAREpins and reversing the fusion clamp. We thus conclude that the cation (Mg/Ca) dependent membrane interaction is a key determinant of the dual clamp/activator function of Synaptotagmin-1.
External linksNat Commun / PubMed:31160571 / PubMed Central
MethodsEM (helical sym.)
Resolution10.4 Å
Structure data

9231

6mti

EMDB-9231: Cryo-EM structure of membrane-bound Synaptotagmin 1 C2AB domains in a complex with the SNAREpin assembly in presence of Mg2+
PDB-6mti: Synaptotagmin-1 C2A, C2B domains and SNARE-pin proteins (5CCI) individually docked into Cryo-EM map of C2AB-SNARE complexes helically organized on lipid nanotube surface in presence of Mg2+
Method: EM (helical sym.) / Resolution: 10.4 Å

Chemicals

ChemComp-MG:
Unknown entry

Source
  • rattus norvegicus (Norway rat)
KeywordsEXOCYTOSIS / SNARE / lipid nanotubes

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