+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-12890 | |||||||||
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Title | Tetanus neurotoxin HC domain in complex with TT104-Fab1 | |||||||||
Map data | Tetanus neurotoxin HC domain in complex with TT104-Fab1 | |||||||||
Sample |
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Function / homology | Function and homology information negative regulation of neurotransmitter secretion / metallopeptidase activity / extracellular region Similarity search - Function | |||||||||
Biological species | Clostridium tetani (bacteria) / Homo sapiens (human) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.9 Å | |||||||||
Authors | Grinzato A / Kandiah E / Zanotti G | |||||||||
Citation | Journal: J Clin Invest / Year: 2021 Title: Exceptionally potent human monoclonal antibodies are effective for prophylaxis and treatment of tetanus in mice. Authors: Marco Pirazzini / Alessandro Grinzato / Davide Corti / Sonia Barbieri / Oneda Leka / Francesca Vallese / Marika Tonellato / Chiara Silacci-Fregni / Luca Piccoli / Eaazhisai Kandiah / ...Authors: Marco Pirazzini / Alessandro Grinzato / Davide Corti / Sonia Barbieri / Oneda Leka / Francesca Vallese / Marika Tonellato / Chiara Silacci-Fregni / Luca Piccoli / Eaazhisai Kandiah / Giampietro Schiavo / Giuseppe Zanotti / Antonio Lanzavecchia / Cesare Montecucco / Abstract: We used human monoclonal antibodies (humAbs) to study the mechanism of neuron intoxication by tetanus neurotoxin and to evaluate these antibodies as a safe preventive and therapeutic substitute for ...We used human monoclonal antibodies (humAbs) to study the mechanism of neuron intoxication by tetanus neurotoxin and to evaluate these antibodies as a safe preventive and therapeutic substitute for hyperimmune sera to treat tetanus in mice. By screening memory B cells from immune donors, we selected 2 tetanus neurotoxin-specific mAbs with exceptionally high neutralizing activities and extensively characterized them both structurally and functionally. We found that these antibodies interfered with the binding and translocation of the neurotoxin into neurons by interacting with 2 epitopes, whose identification pinpoints crucial events in the cellular pathogenesis of tetanus. Our observations explain the neutralization ability of these antibodies, which we found to be exceptionally potent in preventing experimental tetanus when injected into mice long before the toxin. Moreover, their Fab derivatives neutralized tetanus neurotoxin in post-exposure experiments, suggesting their potential for therapeutic use via intrathecal injection. As such, we believe these humAbs, as well as their Fab derivatives, meet the requirements to be considered for prophylactic and therapeutic use in human tetanus and are ready for clinical trials. | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_12890.map.gz | 96.9 MB | EMDB map data format | |
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Header (meta data) | emd-12890-v30.xml emd-12890.xml | 12.1 KB 12.1 KB | Display Display | EMDB header |
Images | emd_12890.png | 48.8 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-12890 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-12890 | HTTPS FTP |
-Validation report
Summary document | emd_12890_validation.pdf.gz | 332.5 KB | Display | EMDB validaton report |
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Full document | emd_12890_full_validation.pdf.gz | 332 KB | Display | |
Data in XML | emd_12890_validation.xml.gz | 6.5 KB | Display | |
Data in CIF | emd_12890_validation.cif.gz | 7.4 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-12890 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-12890 | HTTPS FTP |
-Related structure data
Related structure data | 7oh0MC 7oh1C M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_12890.map.gz / Format: CCP4 / Size: 103 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Tetanus neurotoxin HC domain in complex with TT104-Fab1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.827 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Sample components
-Entire : Tetanus neurotoxin HC domain in complex with TT104-Fab1
Entire | Name: Tetanus neurotoxin HC domain in complex with TT104-Fab1 |
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Components |
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-Supramolecule #1: Tetanus neurotoxin HC domain in complex with TT104-Fab1
Supramolecule | Name: Tetanus neurotoxin HC domain in complex with TT104-Fab1 type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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-Supramolecule #2: Tetanus toxin
Supramolecule | Name: Tetanus toxin / type: complex / ID: 2 / Parent: 1 / Macromolecule list: #1 |
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Source (natural) | Organism: Clostridium tetani (bacteria) |
-Supramolecule #3: FAB TT110
Supramolecule | Name: FAB TT110 / type: complex / ID: 3 / Parent: 1 / Macromolecule list: #2-#3 |
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Source (natural) | Organism: Homo sapiens (human) |
-Macromolecule #1: Tetanus toxin
Macromolecule | Name: Tetanus toxin / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Clostridium tetani (bacteria) |
Molecular weight | Theoretical: 50.476918 KDa |
Sequence | String: EDIDVILKKS TILNLDINND IISDISGFNS SVITYPDAQL VPGINGKAIH LVNNESSEVI VHKAMDIEYN DMFNNFTVSF WLRVPKVSA SHLEQYGTNE YSIISSMKKH SLSIGSGWSV SLKGNNLIWT LKDSAGEVRQ ITFRDLPDKF NAYLANKWVF I TITNDRLS ...String: EDIDVILKKS TILNLDINND IISDISGFNS SVITYPDAQL VPGINGKAIH LVNNESSEVI VHKAMDIEYN DMFNNFTVSF WLRVPKVSA SHLEQYGTNE YSIISSMKKH SLSIGSGWSV SLKGNNLIWT LKDSAGEVRQ ITFRDLPDKF NAYLANKWVF I TITNDRLS SANLYINGVL MGSAEITGLG AIREDNNITL KLDRCNNNNQ YVSIDKFRIF CKALNPKEIE KLYTSYLSIT FL RDFWGNP LRYDTEYYLI PVASSSKDVQ LKNITDYMYL TNAPSYTNGK LNIYYRRLYN GLKFIIKRYT PNNEIDSFVK SGD FIKLYV SYNNNEHIVG YPKDGNAFNN LDRILRVGYN APGIPLYKKM EAVKLRDLKT YSVQLKLYDD KNASLGLVGT HNGQ IGNDP NRDILIASNW YFNHLKDKIL GCDWYFVPTD EGWTND |
-Macromolecule #2: Fab TT104
Macromolecule | Name: Fab TT104 / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 23.647295 KDa |
Sequence | String: QLQLQESGPG LVRPSETLSL TCSVSDDSMG SSTYYWAWIR QPPGKGLEWI GNVYYNGVTY YNSSLESRLT LSIDPSKNQF SLILSSVTA ADTAVYYCAR QADNWFDPWG QGTLVTVSSA STKGPSVFPL APSSKSTSGG TAALGCLVKD YFPEPVTVSW N SGALTSGV ...String: QLQLQESGPG LVRPSETLSL TCSVSDDSMG SSTYYWAWIR QPPGKGLEWI GNVYYNGVTY YNSSLESRLT LSIDPSKNQF SLILSSVTA ADTAVYYCAR QADNWFDPWG QGTLVTVSSA STKGPSVFPL APSSKSTSGG TAALGCLVKD YFPEPVTVSW N SGALTSGV HTFPAVLQSS GLYSLSSVVT VPSSSLGTQT YICNVNHKPS NTKVDKRVEP KSC |
-Macromolecule #3: Fab TT104
Macromolecule | Name: Fab TT104 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 23.462094 KDa |
Sequence | String: IMMTQSPATL SVSPGERATL SCRASQSVGS DLAWYQQKPG QAPRLLIYGA SYRATGIPAR FSGGGSGTEF TLTISSMQSE DFAVYYCQQ YYNWPPYTFG QGTKVEIKRT VAAPSVFIFP PSDEQLKSGT ASVVCLLNNF YPREAKVQWK VDNALQSGNS Q ESVTEQDS ...String: IMMTQSPATL SVSPGERATL SCRASQSVGS DLAWYQQKPG QAPRLLIYGA SYRATGIPAR FSGGGSGTEF TLTISSMQSE DFAVYYCQQ YYNWPPYTFG QGTKVEIKRT VAAPSVFIFP PSDEQLKSGT ASVVCLLNNF YPREAKVQWK VDNALQSGNS Q ESVTEQDS KDSTYSLSST LTLSKADYEK HKVYACEVTH QGLSSPVTKS FNRGEC |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.6 |
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Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 40.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 3.9 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 98170 |
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Initial angle assignment | Type: MAXIMUM LIKELIHOOD |
Final angle assignment | Type: MAXIMUM LIKELIHOOD |