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Yorodumi- EMDB-26456: cryo-EM structures of a synaptobrevin-Munc18-1-syntaxin-1 complex... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-26456 | |||||||||||||||||||||
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Title | cryo-EM structures of a synaptobrevin-Munc18-1-syntaxin-1 complex class1 | |||||||||||||||||||||
Map data | cryo-EM map of a synaptobrevin-Munc18-1-syntaxin-1 complex, conformation 2 | |||||||||||||||||||||
Sample |
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Function / homology | Function and homology information positive regulation of vesicle docking / regulation of acrosomal vesicle exocytosis / positive regulation of glutamate secretion, neurotransmission / negative regulation of SNARE complex assembly / regulation of vesicle fusion / Toxicity of tetanus toxin (tetX) / developmental process involved in reproduction / clathrin-sculpted glutamate transport vesicle membrane / axon target recognition / Toxicity of botulinum toxin type G (botG) ...positive regulation of vesicle docking / regulation of acrosomal vesicle exocytosis / positive regulation of glutamate secretion, neurotransmission / negative regulation of SNARE complex assembly / regulation of vesicle fusion / Toxicity of tetanus toxin (tetX) / developmental process involved in reproduction / clathrin-sculpted glutamate transport vesicle membrane / axon target recognition / Toxicity of botulinum toxin type G (botG) / Toxicity of botulinum toxin type F (botF) / regulation of delayed rectifier potassium channel activity / Toxicity of botulinum toxin type D (botD) / myosin head/neck binding / trans-Golgi Network Vesicle Budding / Other interleukin signaling / Acetylcholine Neurotransmitter Release Cycle / extrinsic component of presynaptic membrane / synaptic vesicle fusion to presynaptic active zone membrane / synaptobrevin 2-SNAP-25-syntaxin-1a-complexin II complex / synaptobrevin 2-SNAP-25-syntaxin-1a complex / synaptobrevin 2-SNAP-25-syntaxin-1a-complexin I complex / clathrin-sculpted monoamine transport vesicle membrane / Toxicity of botulinum toxin type B (botB) / positive regulation of norepinephrine secretion / regulation of synaptic vesicle priming / clathrin-sculpted gamma-aminobutyric acid transport vesicle membrane / Glutamate Neurotransmitter Release Cycle / Norepinephrine Neurotransmitter Release Cycle / zymogen granule membrane / Acetylcholine Neurotransmitter Release Cycle / positive regulation of catecholamine secretion / Serotonin Neurotransmitter Release Cycle / Serotonin Neurotransmitter Release Cycle / GABA synthesis, release, reuptake and degradation / regulated exocytosis / Dopamine Neurotransmitter Release Cycle / GABA synthesis, release, reuptake and degradation / presynaptic dense core vesicle exocytosis / negative regulation of synaptic transmission, GABAergic / synaptic vesicle docking / response to gravity / platelet degranulation / Insertion of tail-anchored proteins into the endoplasmic reticulum membrane / Dopamine Neurotransmitter Release Cycle / neuromuscular synaptic transmission / eosinophil degranulation / Norepinephrine Neurotransmitter Release Cycle / positive regulation of calcium ion-dependent exocytosis / vesicle fusion / synaptic vesicle maturation / vesicle docking / chloride channel inhibitor activity / SNARE complex / Golgi to plasma membrane protein transport / SNAP receptor activity / secretion by cell / presynaptic active zone cytoplasmic component / regulation of vesicle-mediated transport / Glutamate Neurotransmitter Release Cycle / positive regulation of mast cell degranulation / LGI-ADAM interactions / hormone secretion / calcium-ion regulated exocytosis / actomyosin / positive regulation of intracellular protein transport / vesicle docking involved in exocytosis / regulation of exocytosis / neurotransmitter secretion / platelet alpha granule / ATP-dependent protein binding / presynaptic cytosol / neuron projection terminus / protein localization to membrane / neurotransmitter transport / insulin secretion / Sensory processing of sound by inner hair cells of the cochlea / parallel fiber to Purkinje cell synapse / syntaxin-1 binding / SNARE complex assembly / long-term synaptic depression / positive regulation of neurotransmitter secretion / syntaxin binding / synaptic vesicle priming / clathrin-coated vesicle / Lysosome Vesicle Biogenesis / Insertion of tail-anchored proteins into the endoplasmic reticulum membrane / Other interleukin signaling / Golgi Associated Vesicle Biogenesis / myosin binding / Insulin processing / modulation of excitatory postsynaptic potential / exocytosis / positive regulation of exocytosis / synaptic vesicle exocytosis / phospholipase binding / positive regulation of excitatory postsynaptic potential / protein sumoylation / synaptic vesicle endocytosis / endomembrane system Similarity search - Function | |||||||||||||||||||||
Biological species | Rattus norvegicus (Norway rat) | |||||||||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.7 Å | |||||||||||||||||||||
Authors | Rizo J / Bai X / Stepien KP / Xu J / Zhang X | |||||||||||||||||||||
Funding support | United States, 6 items
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Citation | Journal: Sci Adv / Year: 2022 Title: SNARE assembly enlightened by cryo-EM structures of a synaptobrevin-Munc18-1-syntaxin-1 complex. Authors: Karolina P Stepien / Junjie Xu / Xuewu Zhang / Xiao-Chen Bai / Josep Rizo / Abstract: Munc18-1 forms a template to organize assembly of the neuronal SNARE complex that triggers neurotransmitter release, binding first to a closed conformation of syntaxin-1 where its amino-terminal ...Munc18-1 forms a template to organize assembly of the neuronal SNARE complex that triggers neurotransmitter release, binding first to a closed conformation of syntaxin-1 where its amino-terminal region interacts with the SNARE motif, and later binding to synaptobrevin. However, the mechanism of SNARE complex assembly remains unclear. Here, we report two cryo-EM structures of Munc18-1 bound to cross-linked syntaxin-1 and synaptobrevin. The structures allow visualization of how syntaxin-1 opens and reveal how part of the syntaxin-1 amino-terminal region can help nucleate interactions between the amino termini of the syntaxin-1 and synaptobrevin SNARE motifs, while their carboxyl termini bind to distal sites of Munc18-1. These observations, together with mutagenesis, SNARE complex assembly experiments, and fusion assays with reconstituted proteoliposomes, support a model whereby these interactions are critical to initiate SNARE complex assembly and multiple energy barriers enable diverse mechanisms for exquisite regulation of neurotransmitter release. | |||||||||||||||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_26456.map.gz | 14.8 MB | EMDB map data format | |
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Header (meta data) | emd-26456-v30.xml emd-26456.xml | 21.2 KB 21.2 KB | Display Display | EMDB header |
Images | emd_26456.png | 140.6 KB | ||
Others | emd_26456_half_map_1.map.gz emd_26456_half_map_2.map.gz | 17 MB 17 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-26456 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-26456 | HTTPS FTP |
-Related structure data
Related structure data | 7udcMC 7udbC M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_26456.map.gz / Format: CCP4 / Size: 22.2 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||
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Annotation | cryo-EM map of a synaptobrevin-Munc18-1-syntaxin-1 complex, conformation 2 | ||||||||||||||||||||
Voxel size | X=Y=Z: 1.08 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Half map: cryo-EM map of a synaptobrevin-Munc18-1-syntaxin-1 complex, conformation 2,...
File | emd_26456_half_map_1.map | ||||||||||||
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Annotation | cryo-EM map of a synaptobrevin-Munc18-1-syntaxin-1 complex, conformation 2, unfiltered half map 1 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: cryo-EM map of a synaptobrevin-Munc18-1-syntaxin-1 complex, conformation 2,...
File | emd_26456_half_map_2.map | ||||||||||||
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Annotation | cryo-EM map of a synaptobrevin-Munc18-1-syntaxin-1 complex, conformation 2, unfiltered half map 2 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
-Entire : ternary complex of Munc18-1 bound to syntaxin-1A and a SNARE moti...
Entire | Name: ternary complex of Munc18-1 bound to syntaxin-1A and a SNARE motif of synaptobrevin 2 |
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Components |
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-Supramolecule #1: ternary complex of Munc18-1 bound to syntaxin-1A and a SNARE moti...
Supramolecule | Name: ternary complex of Munc18-1 bound to syntaxin-1A and a SNARE motif of synaptobrevin 2 type: complex / Chimera: Yes / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: Rattus norvegicus (Norway rat) |
Recombinant expression | Organism: Escherichia coli BL21(DE3) (bacteria) |
-Macromolecule #1: Syntaxin-binding protein 1
Macromolecule | Name: Syntaxin-binding protein 1 / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Rattus norvegicus (Norway rat) |
Molecular weight | Theoretical: 68.714883 KDa |
Recombinant expression | Organism: Escherichia coli BL21(DE3) (bacteria) |
Sequence | String: GSPGISGGGG GIHMAPIGLK AVVGEKIMHD VIKKVKKKGE WKVLVVDQLS MRMLSSCCKM TDIMTEGITI VEDINKRREP LPSLEAVYL ITPSEKSVHS LISDFKDPPT AKYRAAHVFF TDSCPDALFN ELVKSRAAKV IKTLTEINIA FLPYESQVYS L DSADSFQS ...String: GSPGISGGGG GIHMAPIGLK AVVGEKIMHD VIKKVKKKGE WKVLVVDQLS MRMLSSCCKM TDIMTEGITI VEDINKRREP LPSLEAVYL ITPSEKSVHS LISDFKDPPT AKYRAAHVFF TDSCPDALFN ELVKSRAAKV IKTLTEINIA FLPYESQVYS L DSADSFQS FYSPHKAQMK NPILERLAEQ IATLCATLKE YPAVRYRGEY KDNALLAQLI QDKLDAYKAD DPTMGEGPDK AR SQLLILD RGFDPSSPVL HELTFQAMSY DLLPIENDVY KYETSGIGEA RVKEVLLDED DDLWIALRHK HIAEVSQEVT RSL KDFSSS KRMNTGEKTT MRKLSQMLKK MPQYQKELSK YSTHLHLAED CMKHYQGTVD KLCRVEQDLA MGTDAEGEKI KDPM RAIVP ILLDANVSTY DKIRIILLYI FLKNGITEEN LNKLIQHAQI PPEDSEIITN MAHLGVPIVT DSTLRRRSKP ERKER ISEQ TYQLSRWTPI IKDIMEDTIE DKLDTKHYPY ISTRSSASFS TTAVSARYGH WHKNKAPGEY RSGPRLIIFI LGGVSL NEM RCAYEVTQAN GKWEVLIGST HILTPQKLLD TLKKLNKTDE EISS |
-Macromolecule #2: Syntaxin-1A
Macromolecule | Name: Syntaxin-1A / type: protein_or_peptide / ID: 2 Details: An 8 residue polyalanine stretch was modeled between I149 and A178 of chain B entity Syntaxin-1A. Side chains in this region were not visible and residue numbers are tentative. While ...Details: An 8 residue polyalanine stretch was modeled between I149 and A178 of chain B entity Syntaxin-1A. Side chains in this region were not visible and residue numbers are tentative. While processing in OneDep the polyalanine sequence was updated to 161-TSEEAADM-168 Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Rattus norvegicus (Norway rat) |
Molecular weight | Theoretical: 29.221541 KDa |
Recombinant expression | Organism: Escherichia coli BL21(DE3) (bacteria) |
Sequence | String: GSKDRTQELR TAKDSDDDDD VTVTVDRDRF MDEFFEQVEE IRGFIDKIAE NVEEVKRKHS AILASPNPDE KTKEELEELM SDIKKTANK VRSKLKSIEQ SIEQEEGLNR SSADLRIRKT QHSTLSRKFV EVMSEYNATQ SDYRERAKGR IQRQLEITGR T TTSEEAAD ...String: GSKDRTQELR TAKDSDDDDD VTVTVDRDRF MDEFFEQVEE IRGFIDKIAE NVEEVKRKHS AILASPNPDE KTKEELEELM SDIKKTANK VRSKLKSIEQ SIEQEEGLNR SSADLRIRKT QHSTLSRKFV EVMSEYNATQ SDYRERAKGR IQRQLEITGR T TTSEEAAD MLESGNPAIF ASGIIMDSSI SKQALSEIET RHSEIIKCEN SIRELHDMFM DMAMLVESQG EMIDRIEYNV EH AVDYVER AVSDTKK |
-Macromolecule #3: Synaptobrevin-2
Macromolecule | Name: Synaptobrevin-2 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Rattus norvegicus (Norway rat) |
Molecular weight | Theoretical: 6.645386 KDa |
Recombinant expression | Organism: Escherichia coli BL21(DE3) (bacteria) |
Sequence | String: GSMWNRRLQQ TCAQVDEVVD IMRVNVDKVL ERDQKLSELD DRADALQAGA SQFETSAAK |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 7.5 mg/mL |
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Buffer | pH: 7 |
Grid | Model: Quantifoil R1.2/1.3 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Nominal defocus max: 2.6 µm / Nominal defocus min: 1.6 µm |
Specialist optics | Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Image recording | Film or detector model: GATAN K2 QUANTUM (4k x 4k) / Detector mode: SUPER-RESOLUTION / Number real images: 7401 / Average electron dose: 60.0 e/Å2 |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
Particle selection | Number selected: 5819182 |
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CTF correction | Software - Name: Gctf |
Startup model | Type of model: OTHER / Details: From RELION |
Initial angle assignment | Type: PROJECTION MATCHING / Software - Name: RELION |
Final 3D classification | Number classes: 6 / Software - Name: RELION |
Final angle assignment | Type: PROJECTION MATCHING / Software - Name: RELION |
Final reconstruction | Applied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 3.7 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION / Number images used: 331649 |