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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-25209 | |||||||||
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Title | Structure of human SARS-CoV-2 neutralizing antibody C1C-A3 Fab | |||||||||
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Function / homology | ![]() Maturation of spike protein / viral translation / Translation of Structural Proteins / Virion Assembly and Release / host cell surface / host extracellular space / suppression by virus of host tetherin activity / Induction of Cell-Cell Fusion / structural constituent of virion / host cell endoplasmic reticulum-Golgi intermediate compartment membrane ...Maturation of spike protein / viral translation / Translation of Structural Proteins / Virion Assembly and Release / host cell surface / host extracellular space / suppression by virus of host tetherin activity / Induction of Cell-Cell Fusion / structural constituent of virion / host cell endoplasmic reticulum-Golgi intermediate compartment membrane / entry receptor-mediated virion attachment to host cell / receptor-mediated endocytosis of virus by host cell / Attachment and Entry / membrane fusion / positive regulation of viral entry into host cell / receptor-mediated virion attachment to host cell / receptor ligand activity / host cell surface receptor binding / fusion of virus membrane with host plasma membrane / fusion of virus membrane with host endosome membrane / viral envelope / virion attachment to host cell / SARS-CoV-2 activates/modulates innate and adaptive immune responses / host cell plasma membrane / virion membrane / identical protein binding / membrane / plasma membrane Similarity search - Function | |||||||||
Biological species | ![]() ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 4.3 Å | |||||||||
![]() | Pan J / Abraham J / Yang P / Shankar S | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Structural basis for continued antibody evasion by the SARS-CoV-2 receptor binding domain. Authors: Katherine G Nabel / Sarah A Clark / Sundaresh Shankar / Junhua Pan / Lars E Clark / Pan Yang / Adrian Coscia / Lindsay G A McKay / Haley H Varnum / Vesna Brusic / Nicole V Tolan / Guohai ...Authors: Katherine G Nabel / Sarah A Clark / Sundaresh Shankar / Junhua Pan / Lars E Clark / Pan Yang / Adrian Coscia / Lindsay G A McKay / Haley H Varnum / Vesna Brusic / Nicole V Tolan / Guohai Zhou / Michaël Desjardins / Sarah E Turbett / Sanjat Kanjilal / Amy C Sherman / Anand Dighe / Regina C LaRocque / Edward T Ryan / Casey Tylek / Joel F Cohen-Solal / Anhdao T Darcy / Davide Tavella / Anca Clabbers / Yao Fan / Anthony Griffiths / Ivan R Correia / Jane Seagal / Lindsey R Baden / Richelle C Charles / Jonathan Abraham / ![]() ![]() Abstract: Many studies have examined the impact of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants on neutralizing antibody activity after they have become dominant strains. Here, we ...Many studies have examined the impact of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants on neutralizing antibody activity after they have become dominant strains. Here, we evaluate the consequences of further viral evolution. We demonstrate mechanisms through which the SARS-CoV-2 receptor binding domain (RBD) can tolerate large numbers of simultaneous antibody escape mutations and show that pseudotypes containing up to seven mutations, as opposed to the one to three found in previously studied variants of concern, are more resistant to neutralization by therapeutic antibodies and serum from vaccine recipients. We identify an antibody that binds the RBD core to neutralize pseudotypes for all tested variants but show that the RBD can acquire an N-linked glycan to escape neutralization. Our findings portend continued emergence of escape variants as SARS-CoV-2 adapts to humans. | |||||||||
History |
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Structure visualization
Movie |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 49.2 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 23.9 KB 23.9 KB | Display Display | ![]() |
Images | ![]() | 109.2 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 478.8 KB | Display | ![]() |
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Full document | ![]() | 478.4 KB | Display | |
Data in XML | ![]() | 5.6 KB | Display | |
Data in CIF | ![]() | 6.4 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 7sn2MC ![]() 7sn0C ![]() 7sn1C ![]() 7sn3C M: atomic model generated by this map C: citing same article ( |
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Similar structure data |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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Voxel size | X=Y=Z: 0.825 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
-Entire : SARS-CoV-2_Hexapro in complex with neutralizing antibody C1C-A3 F...
Entire | Name: SARS-CoV-2_Hexapro in complex with neutralizing antibody C1C-A3 Fab from human patient |
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Components |
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-Supramolecule #1: SARS-CoV-2_Hexapro in complex with neutralizing antibody C1C-A3 F...
Supramolecule | Name: SARS-CoV-2_Hexapro in complex with neutralizing antibody C1C-A3 Fab from human patient type: complex / ID: 1 / Parent: 0 / Macromolecule list: all Details: concentration of the SARS-CoV-2 spike protein SARS-CoV-2 Hexapro (trimeric) and human patient antibody C1C-A3 Fab is 0.6 and 0.3 mg/mL in the final complex sample in buffer consisting of 150 ...Details: concentration of the SARS-CoV-2 spike protein SARS-CoV-2 Hexapro (trimeric) and human patient antibody C1C-A3 Fab is 0.6 and 0.3 mg/mL in the final complex sample in buffer consisting of 150 mM NaCl and 25 mM Tris-HCl, pH 7.5. structure deposited here is one receptor binding domain (RBD) in complex with on C1C-A3 Fab determined from the relate wwPDB deposit D_1000258559. |
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Source (natural) | Organism: ![]() ![]() |
Recombinant expression | Organism: ![]() |
Molecular weight | Theoretical: 50 KDa |
-Supramolecule #2: SARS-CoV-2 spike protein SARS-CoV-2 Hexapro
Supramolecule | Name: SARS-CoV-2 spike protein SARS-CoV-2 Hexapro / type: complex / ID: 2 / Parent: 1 / Macromolecule list: #1 |
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Source (natural) | Organism: ![]() ![]() |
Recombinant expression | Organism: ![]() |
-Supramolecule #3: C1C-A3 Fab
Supramolecule | Name: C1C-A3 Fab / type: complex / ID: 3 / Parent: 1 / Macromolecule list: #2-#3 Details: Fab of C1C-A3 antibody recombinantly expressed using sequence sequenced from single B cells from human patient peripheral blood sample |
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Source (natural) | Organism: ![]() |
Recombinant expression | Organism: ![]() |
-Macromolecule #1: Spike glycoprotein
Macromolecule | Name: Spike glycoprotein / type: protein_or_peptide / ID: 1 Details: Severe acute respiratory syndrome coronavirus 2 surface glycoprotein (residues 1-1208) of GeneBank locus QHD43416.1 (amino acids 1-15 are the endogenous signal peptide, 16-1208 are SARS-CoV- ...Details: Severe acute respiratory syndrome coronavirus 2 surface glycoprotein (residues 1-1208) of GeneBank locus QHD43416.1 (amino acids 1-15 are the endogenous signal peptide, 16-1208 are SARS-CoV-2 spike protein residues 16-1208, 1209-1210+1237-1239+1255-1258 are linkers, 1211-1236 are T4 fibritinn foldon, 1240-1254 are the avi-tag for biotinylation, 1259-1265 are the TEV protease cleavage site, 1266-1273 are the 8xHis tag; mutations include R682G, R683S, R685S, F817P, A892P, A899P, A942P, K986P, V987P) Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 141.192203 KDa |
Recombinant expression | Organism: ![]() |
Sequence | String: MFVFLVLLPL VSSQCVNLTT RTQLPPAYTN SFTRGVYYPD KVFRSSVLHS TQDLFLPFFS NVTWFHAIHV SGTNGTKRFD NPVLPFNDG VYFASTEKSN IIRGWIFGTT LDSKTQSLLI VNNATNVVIK VCEFQFCNDP FLGVYYHKNN KSWMESEFRV Y SSANNCTF ...String: MFVFLVLLPL VSSQCVNLTT RTQLPPAYTN SFTRGVYYPD KVFRSSVLHS TQDLFLPFFS NVTWFHAIHV SGTNGTKRFD NPVLPFNDG VYFASTEKSN IIRGWIFGTT LDSKTQSLLI VNNATNVVIK VCEFQFCNDP FLGVYYHKNN KSWMESEFRV Y SSANNCTF EYVSQPFLMD LEGKQGNFKN LREFVFKNID GYFKIYSKHT PINLVRDLPQ GFSALEPLVD LPIGINITRF QT LLALHRS YLTPGDSSSG WTAGAAAYYV GYLQPRTFLL KYNENGTITD AVDCALDPLS ETKCTLKSFT VEKGIYQTSN FRV QPTESI VRFPNITNLC PFGEVFNATR FASVYAWNRK RISNCVADYS VLYNSASFST FKCYGVSPTK LNDLCFTNVY ADSF VIRGD EVRQIAPGQT GKIADYNYKL PDDFTGCVIA WNSNNLDSKV GGNYNYLYRL FRKSNLKPFE RDISTEIYQA GSTPC NGVE GFNCYFPLQS YGFQPTNGVG YQPYRVVVLS FELLHAPATV CGPKKSTNLV KNKCVNFNFN GLTGTGVLTE SNKKFL PFQ QFGRDIADTT DAVRDPQTLE ILDITPCSFG GVSVITPGTN TSNQVAVLYQ DVNCTEVPVA IHADQLTPTW RVYSTGS NV FQTRAGCLIG AEHVNNSYEC DIPIGAGICA SYQTQTNSPG SASSVASQSI IAYTMSLGAE NSVAYSNNSI AIPTNFTI S VTTEILPVSM TKTSVDCTMY ICGDSTECSN LLLQYGSFCT QLNRALTGIA VEQDKNTQEV FAQVKQIYKT PPIKDFGGF NFSQILPDPS KPSKRSPIED LLFNKVTLAD AGFIKQYGDC LGDIAARDLI CAQKFNGLTV LPPLLTDEMI AQYTSALLAG TITSGWTFG AGPALQIPFP MQMAYRFNGI GVTQNVLYEN QKLIANQFNS AIGKIQDSLS STPSALGKLQ DVVNQNAQAL N TLVKQLSS NFGAISSVLN DILSRLDPPE AEVQIDRLIT GRLQSLQTYV TQQLIRAAEI RASANLAATK MSECVLGQSK RV DFCGKGY HLMSFPQSAP HGVVFLHVTY VPAQEKNFTT APAICHDGKA HFPREGVFVS NGTHWFVTQR NFYEPQIITT DNT FVSGNC DVVIGIVNNT VYDPLQPELD SFKEELDKYF KNHTSPDVDL GDISGINASV VNIQKEIDRL NEVAKNLNES LIDL QELGK YEQSGYIPEA PRDGQAYVRK DGEWVLLSTF LSAGGLNDIF EAQKIEWHEG ASGENLYFQG HHHHHHHH |
-Macromolecule #2: neutralizing antibody C1C-A3 Fab heavy chain
Macromolecule | Name: neutralizing antibody C1C-A3 Fab heavy chain / type: protein_or_peptide / ID: 2 Details: amino acids 1-20 are the human tPA signal peptide, 21-22 are a linker, 23-250 are the C1C-A3 Fab heavy chain residues 1-213 (note insertions in CDR loops; numbering should be according to the PDB file). Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 27.022607 KDa |
Recombinant expression | Organism: ![]() |
Sequence | String: MKRGLCCVLL LCGAVFVSPS ASQVQLVESG GGVVQPGRSL RLSCAASGFT FSSYGMHWVR QAPGKGLEWV AVIWYDGTNT YYADSVKGR FTISRDNSKN TLYLQMNSLR AEDTAVYYCA RDLAYRDYVW RYFDLWGRGT LVTVSGASTK GPSVFPLAPS S KSTSGGTA ...String: MKRGLCCVLL LCGAVFVSPS ASQVQLVESG GGVVQPGRSL RLSCAASGFT FSSYGMHWVR QAPGKGLEWV AVIWYDGTNT YYADSVKGR FTISRDNSKN TLYLQMNSLR AEDTAVYYCA RDLAYRDYVW RYFDLWGRGT LVTVSGASTK GPSVFPLAPS S KSTSGGTA ALGCLVKDYF PEPVTVSWNS GALTSGVHTF PAVLQSSGLY SLSSVVTVPS SSLGTQTYIC NVNHKPSNTK VD KRVEPKS CDR |
-Macromolecule #3: neutralizing antibody C1C-A3 Fab light chain
Macromolecule | Name: neutralizing antibody C1C-A3 Fab light chain / type: protein_or_peptide / ID: 3 Details: amino acids 1-20 are the human tPA signal peptide, 21-22 are a linker, 23-238 are the C1C-A3 Fab light chain residues 1-212 (note insertions in CDR loops; numbering should be according to the PDB file). Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() |
Molecular weight | Theoretical: 25.896107 KDa |
Recombinant expression | Organism: ![]() |
Sequence | String: MKRGLCCVLL LCGAVFVSPS ASEIVLTQSP ATLSLSPGER ATLSCRASQS VSTYLAWYQQ KFGQAPRLLI YDASNRATGI PARFSGSGS GTDFTLTISS LEPEDFAVYY CQCRSNWPPG ITFGQGTRLE IKRTVAAPSV FIFPPSDEQL KSGTASVVCL L NNFYPREA ...String: MKRGLCCVLL LCGAVFVSPS ASEIVLTQSP ATLSLSPGER ATLSCRASQS VSTYLAWYQQ KFGQAPRLLI YDASNRATGI PARFSGSGS GTDFTLTISS LEPEDFAVYY CQCRSNWPPG ITFGQGTRLE IKRTVAAPSV FIFPPSDEQL KSGTASVVCL L NNFYPREA KVQWKVDNAL QSGNSQESVT EQDSKDSTYS LSSTLTLSKA DYEKHKVYAC EVTHQGLSSP VTKSFNRGEC |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 0.9 mg/mL | |||||||||
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Buffer | pH: 7.5 Component:
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Grid | Model: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 300 / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Support film - Film thickness: 2.0 nm / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Atmosphere: AIR / Pretreatment - Pressure: 0.039 kPa Details: Quantifoil Cu 1.2/1.3 2nm carbon 300 mesh grids glowed discharge at 15 mA in Pelco EasiGlow | |||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 293 K / Instrument: FEI VITROBOT MARK IV / Details: blot for 4-6 seconds. | |||||||||
Details | this sample was monodisperse. |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Temperature | Min: 77.0 K / Max: 77.0 K |
Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Digitization - Dimensions - Width: 5760 pixel / Digitization - Dimensions - Height: 4092 pixel / Digitization - Sampling interval: 5.0 µm / Number grids imaged: 1 / Number real images: 4583 / Average exposure time: 1.9 sec. / Average electron dose: 56.5 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | C2 aperture diameter: 50.0 µm / Calibrated magnification: 60606 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 2.5 µm / Nominal defocus min: 1.0 µm / Nominal magnification: 105000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |