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- EMDB-70353: Clone 2.1 Fab in complex with chicken IgY CH2 domain (local refin... -
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Open data
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Basic information
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Title | Clone 2.1 Fab in complex with chicken IgY CH2 domain (local refinement) | |||||||||
![]() | sharpened map | |||||||||
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![]() | germinal center evolution / antibody / IMMUNE SYSTEM | |||||||||
Function / homology | ![]() : / Immunoglobulin V-Type / Immunoglobulin V-set domain / Immunoglobulin V-set domain / Immunoglobulin/major histocompatibility complex, conserved site / Immunoglobulins and major histocompatibility complex proteins signature. / Immunoglobulin subtype / Immunoglobulin / Immunoglobulin C-Type / Immunoglobulin C1-set ...: / Immunoglobulin V-Type / Immunoglobulin V-set domain / Immunoglobulin V-set domain / Immunoglobulin/major histocompatibility complex, conserved site / Immunoglobulins and major histocompatibility complex proteins signature. / Immunoglobulin subtype / Immunoglobulin / Immunoglobulin C-Type / Immunoglobulin C1-set / Immunoglobulin C1-set domain / Ig-like domain profile. / Immunoglobulin-like domain / Immunoglobulin-like domain superfamily / Immunoglobulin-like fold Similarity search - Domain/homology | |||||||||
Biological species | ![]() ![]() ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 4.0 Å | |||||||||
![]() | Ozorowski G / Alkutkar T / Ward AB | |||||||||
Funding support | 1 items
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![]() | ![]() Title: Replaying germinal center evolution on a quantified affinity landscape. Authors: William S DeWitt / Ashni A Vora / Tatsuya Araki / Jared G Galloway / Tanwee Alkutkar / Juliana Bortolatto / Tiago B R Castro / Will Dumm / Chris Jennings-Shaffer / Tongqiu Jia / Luka Mesin / ...Authors: William S DeWitt / Ashni A Vora / Tatsuya Araki / Jared G Galloway / Tanwee Alkutkar / Juliana Bortolatto / Tiago B R Castro / Will Dumm / Chris Jennings-Shaffer / Tongqiu Jia / Luka Mesin / Gabriel Ozorowski / Juhee Pae / Duncan K Ralph / Jesse D Bloom / Armita Nourmohammad / Yun S Song / Andrew B Ward / Tyler N Starr / Frederick A Matsen / Gabriel D Victora / ![]() Abstract: Darwinian evolution of immunoglobulin genes within germinal centers (GC) underlies the progressive increase in antibody affinity following antigen exposure. Whereas the mechanics of how competition ...Darwinian evolution of immunoglobulin genes within germinal centers (GC) underlies the progressive increase in antibody affinity following antigen exposure. Whereas the mechanics of how competition between GC B cells drives increased affinity are well established, the dynamical evolutionary features of this process remain poorly characterized. We devised an experimental evolution model in which we "replay" over one hundred instances of a clonally homogenous GC reaction and follow the selective process by assigning affinities to all cells using deep mutational scanning. Our data reveal how GCs achieve predictable evolutionary outcomes through the cumulative effects of many rounds of imperfect selection, acting on a landscape shaped heavily by somatic hypermutation (SHM) targeting biases. Using time-calibrated models, we show that apparent features of GC evolution such as permissiveness to low-affinity lineages and early plateauing of affinity are best explained by survivorship biases that distort our view of how affinity progresses over time. | |||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 59.6 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 20.3 KB 20.3 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 8.5 KB | Display | ![]() |
Images | ![]() | 56.8 KB | ||
Masks | ![]() | 64 MB | ![]() | |
Filedesc metadata | ![]() | 6.8 KB | ||
Others | ![]() ![]() | 59.3 MB 59.3 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 787.3 KB | Display | ![]() |
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Full document | ![]() | 786.9 KB | Display | |
Data in XML | ![]() | 16.4 KB | Display | |
Data in CIF | ![]() | 21.1 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 9odbMC M: atomic model generated by this map C: citing same article ( |
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Similar structure data | Similarity search - Function & homology ![]() |
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Links
EMDB pages | ![]() ![]() |
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Map
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Annotation | sharpened map | ||||||||||||||||||||||||||||||||||||
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
-Mask #1
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Projections & Slices |
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Density Histograms |
-Half map: half map A
File | emd_70353_half_map_1.map | ||||||||||||
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Annotation | half map A | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: half map B
File | emd_70353_half_map_2.map | ||||||||||||
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Annotation | half map B | ||||||||||||
Projections & Slices |
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Density Histograms |
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Sample components
-Entire : Clone 2.1 Fab in complex with chicken IgY CH2 domain
Entire | Name: Clone 2.1 Fab in complex with chicken IgY CH2 domain |
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Components |
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-Supramolecule #1: Clone 2.1 Fab in complex with chicken IgY CH2 domain
Supramolecule | Name: Clone 2.1 Fab in complex with chicken IgY CH2 domain / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#3 |
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Source (natural) | Organism: ![]() ![]() |
-Macromolecule #1: chicken IgY heavy chain,Ig-like domain-containing protein
Macromolecule | Name: chicken IgY heavy chain,Ig-like domain-containing protein type: protein_or_peptide / ID: 1 / Number of copies: 2 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 59.474793 KDa |
Recombinant expression | Organism: ![]() |
Sequence | String: AVTLDESGGG LQTPGGTLSL VCEGSGFTFS SFHMQWVRQA LGKGLEWVAG ISGSGKYTSY GAAVKGRATI SRDNGQSTVR LQLNNLKAE DTGTYYCTRS SCGGCCDWSP YTTGCGAYST GTIDAWGHGT EVIVSSASPT SPPRLYPLSA CCSDSAVPPA V GCLLSPSS ...String: AVTLDESGGG LQTPGGTLSL VCEGSGFTFS SFHMQWVRQA LGKGLEWVAG ISGSGKYTSY GAAVKGRATI SRDNGQSTVR LQLNNLKAE DTGTYYCTRS SCGGCCDWSP YTTGCGAYST GTIDAWGHGT EVIVSSASPT SPPRLYPLSA CCSDSAVPPA V GCLLSPSS AGGISWEGSG GTAVAGRVSG TPVKLSFVRL SPGEKRKSFV CSAAPGGALL KKEVQVCRVD PVPPVAPEVQ VL HPSSCTP SQSESVELLC LVTGFSPASA EVEWLVDGVG GLLVASQSPA VRSGSTYSLS SRVNVSGTDW REGKSYSCRV RHP ATNTVV EDHVKGCPDG AQSCSPIQLY AIPPSPGELY ISLDAKLRCL VVNLPSDSSL SVTWTREKSG NLRPDPMVLQ EHFN GTYSA SSAVPVSTQD WLSGERFTCT VQHEELPLPL SKSVYRNTGP TTPPLIYPFA PHPEELSLSR VTLSCLVRGF RPRDI EIRW LRDHRAVPAT EFVTTAVLPE ERTANGAGGD GDTFFVYSKM SVETAKWNGG TVFACMAVHE ALPMRFSQRT LQKQAG K UniProtKB: Ig-like domain-containing protein |
-Macromolecule #2: Clone 2.1 Fab heavy chain
Macromolecule | Name: Clone 2.1 Fab heavy chain / type: protein_or_peptide / ID: 2 / Number of copies: 2 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 25.972678 KDa |
Recombinant expression | Organism: ![]() |
Sequence | String: EVQLQESGPS LVKPSQTLSL TCSVTGASIT SGYWNWIRKF PGNRLEYMGY ISYSGGTYYN PSLKSRISIT RDTSKNQYYL QLNSVTTED TATYYCARDF DVWGAGTTVT VSSASTKGPS VFPLAPSSKS TSGGTAALGC LVKDYFPEPV TVSWNSGALT S GVHTFPAV ...String: EVQLQESGPS LVKPSQTLSL TCSVTGASIT SGYWNWIRKF PGNRLEYMGY ISYSGGTYYN PSLKSRISIT RDTSKNQYYL QLNSVTTED TATYYCARDF DVWGAGTTVT VSSASTKGPS VFPLAPSSKS TSGGTAALGC LVKDYFPEPV TVSWNSGALT S GVHTFPAV LQSSGLYSLS SVVTVPSSSL GTQTYICNVN HKPSNTKVDK RVEPKSCDKT HTSRHHHHHH SGSDYKDDDD K |
-Macromolecule #3: Clone 2.1 Fab light chain
Macromolecule | Name: Clone 2.1 Fab light chain / type: protein_or_peptide / ID: 3 / Number of copies: 2 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 23.56107 KDa |
Recombinant expression | Organism: ![]() |
Sequence | String: DIVMTQSQKF MSTSVGDRVS VTCKASQNVG TNVGWFQQKP GQSPKSLIYS ASYRYSGVPD RFTGSGSGTD FTLTISNVQS EDLAEYFCH QYYSYPLTFG SGTKLEIKRT VAAPSVFIFP PSDEQLKSGT ASVVCLLNNF YPREAKVQWK VDNALQSGNS Q ESVTEQDS ...String: DIVMTQSQKF MSTSVGDRVS VTCKASQNVG TNVGWFQQKP GQSPKSLIYS ASYRYSGVPD RFTGSGSGTD FTLTISNVQS EDLAEYFCH QYYSYPLTFG SGTKLEIKRT VAAPSVFIFP PSDEQLKSGT ASVVCLLNNF YPREAKVQWK VDNALQSGNS Q ESVTEQDS KDSTYSLSST LTLSKADYEK HKVYACEVTH QGLSSPVTKS FNRGEC |
-Macromolecule #4: 2-acetamido-2-deoxy-beta-D-glucopyranose
Macromolecule | Name: 2-acetamido-2-deoxy-beta-D-glucopyranose / type: ligand / ID: 4 / Number of copies: 2 / Formula: NAG |
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Molecular weight | Theoretical: 221.208 Da |
Chemical component information | ![]() ChemComp-NAG: |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 1 mg/mL |
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Buffer | pH: 7.4 |
Grid | Model: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 300 / 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 TALOS ARCTICA |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Average electron dose: 50.0 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.5 µm / Nominal defocus min: 0.8 µm / Nominal magnification: 36000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | ![]() Model: Talos Arctica / Image courtesy: FEI Company |