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Yorodumi- EMDB-26456: cryo-EM structures of a synaptobrevin-Munc18-1-syntaxin-1 complex... -
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Basic information
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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 | |||||||||||||||||||||
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Keywords | Munc18 / syntaxin / synaptobrevin / SNARE / membrane fusion / neurotransmitter release / EXOCYTOSIS | |||||||||||||||||||||
| Function / homology | Function and homology informationpositive regulation of vesicle docking / regulation of acrosomal vesicle exocytosis / positive regulation of glutamate secretion, neurotransmission / negative regulation of SNARE complex assembly / axon target recognition / Toxicity of tetanus toxin (tetX) / developmental process involved in reproduction / Toxicity of botulinum toxin type G (botG) / clathrin-sculpted glutamate transport vesicle membrane / regulation of vesicle fusion ...positive regulation of vesicle docking / regulation of acrosomal vesicle exocytosis / positive regulation of glutamate secretion, neurotransmission / negative regulation of SNARE complex assembly / axon target recognition / Toxicity of tetanus toxin (tetX) / developmental process involved in reproduction / Toxicity of botulinum toxin type G (botG) / clathrin-sculpted glutamate transport vesicle membrane / regulation of vesicle fusion / regulation of delayed rectifier potassium channel activity / Toxicity of botulinum toxin type F (botF) / Toxicity of botulinum toxin type D (botD) / trans-Golgi Network Vesicle Budding / Toxicity of botulinum toxin type B (botB) / negative regulation of synaptic transmission, GABAergic / clathrin-sculpted gamma-aminobutyric acid transport vesicle membrane / myosin head/neck binding / GABA synthesis, release, reuptake and degradation / zymogen granule membrane / Acetylcholine Neurotransmitter Release Cycle / Other interleukin signaling / synaptobrevin 2-SNAP-25-syntaxin-1a-complexin II complex / synaptobrevin 2-SNAP-25-syntaxin-1a complex / platelet degranulation / presynaptic dense core vesicle exocytosis / synaptobrevin 2-SNAP-25-syntaxin-1a-complexin I complex / clathrin-sculpted monoamine transport vesicle membrane / extrinsic component of presynaptic membrane / Glutamate Neurotransmitter Release Cycle / Norepinephrine Neurotransmitter Release Cycle / synaptic vesicle maturation / Acetylcholine Neurotransmitter Release Cycle / Serotonin Neurotransmitter Release Cycle / Serotonin Neurotransmitter Release Cycle / GABA synthesis, release, reuptake and degradation / positive regulation of norepinephrine secretion / positive regulation of catecholamine secretion / Dopamine Neurotransmitter Release Cycle / synaptic vesicle docking / eosinophil degranulation / regulation of synaptic vesicle priming / regulated exocytosis / presynaptic active zone cytoplasmic component / Insertion of tail-anchored proteins into the endoplasmic reticulum membrane / positive regulation of intracellular protein transport / Dopamine Neurotransmitter Release Cycle / Norepinephrine Neurotransmitter Release Cycle / positive regulation of calcium ion-dependent exocytosis / regulation of vesicle-mediated transport / positive regulation of mast cell degranulation / vesicle docking / regulation of exocytosis / secretion by cell / chloride channel inhibitor activity / Glutamate Neurotransmitter Release Cycle / platelet alpha granule / SNARE complex / SNAP receptor activity / calcium-ion regulated exocytosis / neuromuscular synaptic transmission / vesicle fusion / actomyosin / hormone secretion / LGI-ADAM interactions / neuron projection terminus / Golgi to plasma membrane protein transport / ATP-dependent protein binding / neurotransmitter secretion / vesicle docking involved in exocytosis / long-term synaptic depression / protein localization to membrane / clathrin-coated vesicle / syntaxin binding / insulin secretion / syntaxin-1 binding / Sensory processing of sound by inner hair cells of the cochlea / Other interleukin signaling / Insertion of tail-anchored proteins into the endoplasmic reticulum membrane / Lysosome Vesicle Biogenesis / SNARE complex assembly / positive regulation of neurotransmitter secretion / neurotransmitter transport / Golgi Associated Vesicle Biogenesis / Insulin processing / myosin binding / synaptic vesicle priming / parallel fiber to Purkinje cell synapse / response to gravity / exocytosis / modulation of excitatory postsynaptic potential / positive regulation of exocytosis / synaptic vesicle exocytosis / protein sumoylation / synaptic vesicle endocytosis / positive regulation of excitatory postsynaptic potential / response to glucose / negative regulation of protein-containing complex assembly / calcium channel inhibitor activity / phospholipase binding Similarity search - Function | |||||||||||||||||||||
| Biological species | ![]() | |||||||||||||||||||||
| 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: 2022Title: 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
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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 | 24.2 KB 24.2 KB | Display Display | EMDB header |
| Images | emd_26456.png | 140.6 KB | ||
| Filedesc metadata | emd-26456.cif.gz | 7.4 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 |
-Validation report
| Summary document | emd_26456_validation.pdf.gz | 727.6 KB | Display | EMDB validaton report |
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| Full document | emd_26456_full_validation.pdf.gz | 727.2 KB | Display | |
| Data in XML | emd_26456_validation.xml.gz | 10.1 KB | Display | |
| Data in CIF | emd_26456_validation.cif.gz | 11.8 KB | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-26456 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-26456 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 7udcMC ![]() 7udbC M: atomic model generated by this map C: citing same article ( |
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| Similar structure data | Similarity search - Function & homology F&H Search |
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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| Related items in Molecule of the Month |
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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 | ||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
| 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 |
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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 / ID: 1 / Parent: 0 / Macromolecule list: all |
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| Source (natural) | Organism: ![]() |
-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: ![]() |
| Molecular weight | Theoretical: 68.714883 KDa |
| Recombinant expression | Organism: ![]() |
| 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 UniProtKB: Syntaxin-binding protein 1 |
-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: ![]() |
| Molecular weight | Theoretical: 29.221541 KDa |
| Recombinant expression | Organism: ![]() |
| 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 UniProtKB: Syntaxin-1A |
-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: ![]() |
| Molecular weight | Theoretical: 6.645386 KDa |
| Recombinant expression | Organism: ![]() |
| Sequence | String: GSMWNRRLQQ TCAQVDEVVD IMRVNVDKVL ERDQKLSELD DRADALQAGA SQFETSAAK UniProtKB: Vesicle-associated membrane protein 2 |
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | particle |
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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 |
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Electron microscopy
| Microscope | FEI TITAN KRIOS |
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| Specialist optics | Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV |
| 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 |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.6 µm / Nominal defocus min: 1.6 µm |
| Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Keywords
Authors
United States, 6 items
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Processing
FIELD EMISSION GUN
