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Yorodumi- EMDB-27786: Cryo-EM structure of 334 Fab in complex with recombinant shortene... -
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-Basic information
Entry | Database: EMDB / ID: EMD-27786 | |||||||||
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Title | Cryo-EM structure of 334 Fab in complex with recombinant shortened Plasmodium falciparum circumsporozoite protein (rsCSP) | |||||||||
Map data | Cryo-EM structure of 334 Fab in complex with recombinant shortened Plasmodium falciparum circumsporozoite protein (rsCSP) | |||||||||
Sample |
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Keywords | malaria antibody / PfCSP / IMMUNE SYSTEM | |||||||||
Biological species | Plasmodium falciparum (malaria parasite P. falciparum) / Homo sapiens (human) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.62 Å | |||||||||
Authors | Martin GM / Ward AB | |||||||||
Funding support | United States, 1 items
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Citation | Journal: Nat Commun / Year: 2023 Title: Affinity-matured homotypic interactions induce spectrum of PfCSP structures that influence protection from malaria infection. Authors: Gregory M Martin / Jonathan L Torres / Tossapol Pholcharee / David Oyen / Yevel Flores-Garcia / Grace Gibson / Re'em Moskovitz / Nathan Beutler / Diana D Jung / Jeffrey Copps / Wen-Hsin Lee ...Authors: Gregory M Martin / Jonathan L Torres / Tossapol Pholcharee / David Oyen / Yevel Flores-Garcia / Grace Gibson / Re'em Moskovitz / Nathan Beutler / Diana D Jung / Jeffrey Copps / Wen-Hsin Lee / Gonzalo Gonzalez-Paez / Daniel Emerling / Randall S MacGill / Emily Locke / C Richter King / Fidel Zavala / Ian A Wilson / Andrew B Ward / Abstract: The generation of high-quality antibody responses to Plasmodium falciparum (Pf) circumsporozoite protein (PfCSP), the primary surface antigen of Pf sporozoites, is paramount to the development of an ...The generation of high-quality antibody responses to Plasmodium falciparum (Pf) circumsporozoite protein (PfCSP), the primary surface antigen of Pf sporozoites, is paramount to the development of an effective malaria vaccine. Here we present an in-depth structural and functional analysis of a panel of potent antibodies encoded by the immunoglobulin heavy chain variable (IGHV) gene IGHV3-33, which is among the most prevalent and potent antibody families induced in the anti-PfCSP immune response and targets the Asn-Ala-Asn-Pro (NANP) repeat region. Cryo-electron microscopy (cryo-EM) reveals a remarkable spectrum of helical antibody-PfCSP structures stabilized by homotypic interactions between tightly packed fragments antigen binding (Fabs), many of which correlate with somatic hypermutation. We demonstrate a key role of these mutated homotypic contacts for high avidity binding to PfCSP and in protection from Pf malaria infection. Together, these data emphasize the importance of anti-homotypic affinity maturation in the frequent selection of IGHV3-33 antibodies and highlight key features underlying the potent protection of this antibody family. | |||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_27786.map.gz | 37.8 MB | EMDB map data format | |
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Header (meta data) | emd-27786-v30.xml emd-27786.xml | 17.8 KB 17.8 KB | Display Display | EMDB header |
Images | emd_27786.png | 126.2 KB | ||
Filedesc metadata | emd-27786.cif.gz | 5.8 KB | ||
Others | emd_27786_half_map_1.map.gz emd_27786_half_map_2.map.gz | 37.3 MB 37.3 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-27786 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-27786 | HTTPS FTP |
-Validation report
Summary document | emd_27786_validation.pdf.gz | 842.2 KB | Display | EMDB validaton report |
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Full document | emd_27786_full_validation.pdf.gz | 841.8 KB | Display | |
Data in XML | emd_27786_validation.xml.gz | 11.5 KB | Display | |
Data in CIF | emd_27786_validation.cif.gz | 13.3 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-27786 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-27786 | HTTPS FTP |
-Related structure data
Related structure data | 8dyyMC 8dytC 8dywC 8dyxC 8dz3C 8dz4C 8dz5C 8ekfC M: atomic model generated by this map C: citing same article (ref.) |
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-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_27786.map.gz / Format: CCP4 / Size: 40.6 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Annotation | Cryo-EM structure of 334 Fab in complex with recombinant shortened Plasmodium falciparum circumsporozoite protein (rsCSP) | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.15 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Half map: Cryo-EM structure of 334 Fab in complex with...
File | emd_27786_half_map_1.map | ||||||||||||
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Annotation | Cryo-EM structure of 334 Fab in complex with recombinant shortened Plasmodium falciparum circumsporozoite protein (rsCSP) | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Cryo-EM structure of 334 Fab in complex with...
File | emd_27786_half_map_2.map | ||||||||||||
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Annotation | Cryo-EM structure of 334 Fab in complex with recombinant shortened Plasmodium falciparum circumsporozoite protein (rsCSP) | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
-Entire : 334 Fab - rsCSP complex
Entire | Name: 334 Fab - rsCSP complex |
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Components |
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-Supramolecule #1: 334 Fab - rsCSP complex
Supramolecule | Name: 334 Fab - rsCSP complex / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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-Supramolecule #2: Circumsporozoite protein
Supramolecule | Name: Circumsporozoite protein / type: complex / ID: 2 / Parent: 1 / Macromolecule list: #1 |
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Source (natural) | Organism: Plasmodium falciparum (malaria parasite P. falciparum) |
-Supramolecule #3: 334 Fab heavy chain, 334 Fab light chain
Supramolecule | Name: 334 Fab heavy chain, 334 Fab light chain / type: complex / ID: 3 / Parent: 1 / Macromolecule list: #2-#3 |
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Source (natural) | Organism: Homo sapiens (human) |
-Macromolecule #1: Circumsporozoite protein
Macromolecule | Name: Circumsporozoite protein / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Plasmodium falciparum (malaria parasite P. falciparum) |
Molecular weight | Theoretical: 30.232156 KDa |
Recombinant expression | Organism: Escherichia coli (E. coli) |
Sequence | String: YGSSSNTRVL NELNYDNAGT NLYNELEMNY YGKQENWYSL KKNSRSLGEN DDGNNEDNEK LRKPKHKKLK QPADGNPDPN ANPNVDPNA NPNVDPNANP NVDPNANPNA NPNANPNANP NANPNANPNA NPNANPNANP NANPNANPNA NPNANPNANP N ANPNANPN ...String: YGSSSNTRVL NELNYDNAGT NLYNELEMNY YGKQENWYSL KKNSRSLGEN DDGNNEDNEK LRKPKHKKLK QPADGNPDPN ANPNVDPNA NPNVDPNANP NVDPNANPNA NPNANPNANP NANPNANPNA NPNANPNANP NANPNANPNA NPNANPNANP N ANPNANPN KNNQGNGQGH NMPNDPNRNV DENANANSAV KNNNNEEPSD KHIKEYLNKI QNSLSTEWSP CSVTCGNGIQ VR IKPGSAN KPKDELDYAN DIEKKICKME KCSSVFNVVN S |
-Macromolecule #2: 334 Fab heavy chain
Macromolecule | Name: 334 Fab heavy chain / type: protein_or_peptide / ID: 2 / Number of copies: 9 / Enantiomer: LEVO |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 24.292213 KDa |
Recombinant expression | Organism: Cricetulus griseus (Chinese hamster) |
Sequence | String: QVQLVESGGG VVQPGRSLTL SCAASGFTFS NYGMHWVRQT PGKGLAWVAI IWYDGSKTYY EDSVKGRFTI SRDNSKNTLY LQMNSLRVD DTAVYYCARV RHSSSRHGSA FDLWGQGTLV TVSSASTKGP SVFPLAPSSK STSGGTAALG CLVKDYFPEP V TVSWNSGA ...String: QVQLVESGGG VVQPGRSLTL SCAASGFTFS NYGMHWVRQT PGKGLAWVAI IWYDGSKTYY EDSVKGRFTI SRDNSKNTLY LQMNSLRVD DTAVYYCARV RHSSSRHGSA FDLWGQGTLV TVSSASTKGP SVFPLAPSSK STSGGTAALG CLVKDYFPEP V TVSWNSGA LTSGVHTFPA VLQSSGLYSL SSVVTVPSSS LGTQTYICNV NHKPSNTKVD KKVEPKSCD |
-Macromolecule #3: 334 Fab light chain
Macromolecule | Name: 334 Fab light chain / type: protein_or_peptide / ID: 3 / Number of copies: 9 / Enantiomer: LEVO |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 23.621119 KDa |
Recombinant expression | Organism: Cricetulus griseus (Chinese hamster) |
Sequence | String: VIQMTQSPST LSASVGDRVT ITCRASQSVS TWLAWYQQKP GQGPKLLIYE ASSLESGVPS RFSGSGSGTE FTLTISSLQP DDFATYYCQ QYNSYSFWTF GQGTKVEIKR TVAAPSVFIF PPSDEQLKSG TASVVCLLNN FYPREAKVQW KVDNALQSGN S QESVTEQD ...String: VIQMTQSPST LSASVGDRVT ITCRASQSVS TWLAWYQQKP GQGPKLLIYE ASSLESGVPS RFSGSGSGTE FTLTISSLQP DDFATYYCQ QYNSYSFWTF GQGTKVEIKR TVAAPSVFIF PPSDEQLKSG TASVVCLLNN FYPREAKVQW KVDNALQSGN S QESVTEQD SKDSTYSLSS TLTLSKADYE KHKVYACEVT HQGLSSPVTK SFNRGEC |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.4 |
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Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | FEI TALOS ARCTICA |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 50.0 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 0.2 µm / Nominal defocus min: 0.5 µm |
Experimental equipment | Model: Talos Arctica / Image courtesy: FEI Company |
-Image processing
Startup model | Type of model: NONE |
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Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 3.62 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 458706 |
Initial angle assignment | Type: PROJECTION MATCHING |
Final angle assignment | Type: PROJECTION MATCHING |