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- EMDB-27443: Cryo-EM structure of influenza A virus A/Bayern/7/1995 hemaggluti... -

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Basic information

Entry
Database: EMDB / ID: EMD-27443
TitleCryo-EM structure of influenza A virus A/Bayern/7/1995 hemagglutinin bound to CR6261 Fab
Map data
Sample
  • Complex: Influenza A virus A/Bayern/7/1995 hemagglutinin bound to CR6261 Fab
    • Protein or peptide: hemagglutinin
    • Protein or peptide: CR6261 Fab heavy chain
    • Protein or peptide: CR6261 Fab light chain
  • Ligand: 2-acetamido-2-deoxy-beta-D-glucopyranose
  • Ligand: water
Biological speciesInfluenza A virus / Homo sapiens (human)
Methodsingle particle reconstruction / cryo EM / Resolution: 2.61 Å
AuthorsSeattle Structural Genomics Center for Infectious Disease (SSGCID)
Funding support United States, 1 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)HHSN272201700059C United States
CitationJournal: To be published
Title: CryoEM structure of influenza A virus A/Bayern/7/1995 hemagglutinin bound to CR6261 Fab
Authors: Yang M / Dranow DM / Edwards TE / Horanyi PS / Lorimer DD
History
DepositionJun 29, 2022-
Header (metadata) releaseDec 14, 2022-
Map releaseDec 14, 2022-
UpdateDec 14, 2022-
Current statusDec 14, 2022Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_27443.png

Downloads & links

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Map

FileDownload / File: emd_27443.map.gz / Format: CCP4 / Size: 662.8 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Voxel sizeX=Y=Z: 0.675 Å
Density
Contour LevelBy AUTHOR: 10.0
Minimum - Maximum-38.581264 - 57.94881
Average (Standard dev.)4.4558687e-13 (±1.0)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderZYX
Origin000
Dimensions558558558
Spacing558558558
CellA=B=C: 376.65 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: #1

Fileemd_27443_half_map_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: #2

Fileemd_27443_half_map_2.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Sample components

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Entire : Influenza A virus A/Bayern/7/1995 hemagglutinin bound to CR6261 Fab

EntireName: Influenza A virus A/Bayern/7/1995 hemagglutinin bound to CR6261 Fab
Components
  • Complex: Influenza A virus A/Bayern/7/1995 hemagglutinin bound to CR6261 Fab
    • Protein or peptide: hemagglutinin
    • Protein or peptide: CR6261 Fab heavy chain
    • Protein or peptide: CR6261 Fab light chain
  • Ligand: 2-acetamido-2-deoxy-beta-D-glucopyranose
  • Ligand: water

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Supramolecule #1: Influenza A virus A/Bayern/7/1995 hemagglutinin bound to CR6261 Fab

SupramoleculeName: Influenza A virus A/Bayern/7/1995 hemagglutinin bound to CR6261 Fab
type: complex / Chimera: Yes / ID: 1 / Parent: 0 / Macromolecule list: #1-#3
Molecular weightTheoretical: 450 KDa

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Macromolecule #1: hemagglutinin

MacromoleculeName: hemagglutinin / type: protein_or_peptide / ID: 1 / Number of copies: 3 / Enantiomer: LEVO
Source (natural)Organism: Influenza A virus / Strain: A/Bayern/7/1995(H1N1)
Molecular weightTheoretical: 64.569223 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MVLVNQSHQG FNKEHTSKMV SAIVLYVLLA AAAHSAFAGS DTICIGYHAN NSTDTVDTVL EKNVTVTHSV NLLEDSHNGK LCRLKGTAP LQLGNCSVAG WILGNPECES LFSKESWSYI AETPKPENGT CYPGYFADYE ELREQLSSVS SFERFEIFPK E SSWPNHTV ...String:
MVLVNQSHQG FNKEHTSKMV SAIVLYVLLA AAAHSAFAGS DTICIGYHAN NSTDTVDTVL EKNVTVTHSV NLLEDSHNGK LCRLKGTAP LQLGNCSVAG WILGNPECES LFSKESWSYI AETPKPENGT CYPGYFADYE ELREQLSSVS SFERFEIFPK E SSWPNHTV TKGVTAACSH NGKSSFYKNL LWLTEKNGLY PNLSKSYVNN KEKEVLVLWG VHHPSNIGDQ RAIYHTENAY VS VVSSHYS RRFTPEIAKR PKVRGQEGRI NYYWTLLEPG DTIIFEANGN LIAPWYAFAL SRGFGSGIIT SNASMGECDA KCQ TPQGAI NSSLPFQNVH PVTIGECPKY VRSTKLRMVT GLRNIPSIQS RGLFGAIAGF IEGGWTGMID GWYGYHHQNE QGSG YAADQ KSTQNAINGI TNKVNSVIEK MNTQFTAVGK EFNKLERRME NLNKKVDDGF LDIWTYNAEL LVLLENERTL DFHDS NVKN LYEKVKNQLK NNAKEIGNGC FEFYHKCNNE CMESVKNGTY DYPKYSEEFL VPRGSPGSGY IPEAPRDGQA YVRKDG EWV LLSTFLGHHH HHH

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Macromolecule #2: CR6261 Fab heavy chain

MacromoleculeName: CR6261 Fab heavy chain / type: protein_or_peptide / ID: 2 / Number of copies: 3 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 27.066621 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: MEWSWVFLFF LSVTTGVHSE VQLVESGAEV KKPGSSVKVS CKASGGPFRS YAISWVRQAP GQGPEWMGGI IPIFGTTKYA PKFQGRVTI TADDFAGTVY MELSSLRSED TAMYYCAKHM GYQVRETMDV WGKGTTVTVS SASTKGPSVF PLAPSSKSTS G GTAALGCL ...String:
MEWSWVFLFF LSVTTGVHSE VQLVESGAEV KKPGSSVKVS CKASGGPFRS YAISWVRQAP GQGPEWMGGI IPIFGTTKYA PKFQGRVTI TADDFAGTVY MELSSLRSED TAMYYCAKHM GYQVRETMDV WGKGTTVTVS SASTKGPSVF PLAPSSKSTS G GTAALGCL VKDYFPEPVT VSWNSGALTS GVHTFPAVLQ SSGLYSLSSV VTVPSSSLGT QTYICNVNHK PSNTKVDKRV EP KSCDKHH HHHH

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Macromolecule #3: CR6261 Fab light chain

MacromoleculeName: CR6261 Fab light chain / type: protein_or_peptide / ID: 3 / Number of copies: 3 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 25.514395 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: MEWSWVFLFF LSVTTGVHSQ SVLTQPPSVS AAPGQKVTIS CSGSSSNIGN DYVSWYQQLP GTAPKLLIYD NNKRPSGIPD RFSGSKSGT SATLGITGLQ TGDEANYYCA TWDRRPTAYV VFGGGTKLTV LGAAAGQPKA APSVTLFPPS SEELQANKAT L VCLISDFY ...String:
MEWSWVFLFF LSVTTGVHSQ SVLTQPPSVS AAPGQKVTIS CSGSSSNIGN DYVSWYQQLP GTAPKLLIYD NNKRPSGIPD RFSGSKSGT SATLGITGLQ TGDEANYYCA TWDRRPTAYV VFGGGTKLTV LGAAAGQPKA APSVTLFPPS SEELQANKAT L VCLISDFY PGAVTVAWKA DSSPVKAGVE TTTPSKQSNN KYAASSYLSL TPEQWKSHRS YSCQVTHEGS TVEKTVAPTE CS

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Macromolecule #6: 2-acetamido-2-deoxy-beta-D-glucopyranose

MacromoleculeName: 2-acetamido-2-deoxy-beta-D-glucopyranose / type: ligand / ID: 6 / Number of copies: 21 / Formula: NAG
Molecular weightTheoretical: 221.208 Da
Chemical component information

ChemComp-NAG:
2-acetamido-2-deoxy-beta-D-glucopyranose / N-Acetylglucosamine

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Macromolecule #7: water

MacromoleculeName: water / type: ligand / ID: 7 / Number of copies: 140 / Formula: HOH
Molecular weightTheoretical: 18.015 Da
Chemical component information

ChemComp-HOH:
WATER / Water

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

Concentration0.5 mg/mL
BufferpH: 8
Component:
ConcentrationFormulaName
150.0 mMNaClSodium chlorideSodium Chloride
25.0 mMTris(hydroxymethyl)aminomethane hydrochloride
GridModel: EMS Lacey Carbon / Material: COPPER / Support film - Material: GRAPHENE OXIDE / Support film - topology: LACEY
VitrificationCryogen name: ETHANE / Chamber humidity: 90 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Cs: 2.7 mm / Nominal defocus max: 2.5 µm / Nominal defocus min: 1.0 µm / Nominal magnification: 130000
Sample stageCooling holder cryogen: NITROGEN
Image recordingFilm or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Number grids imaged: 1 / Number real images: 12095 / Average exposure time: 2.307 sec. / Average electron dose: 80.0 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Startup modelType of model: PDB ENTRY
PDB model - PDB ID:

6uyn

Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: cryoSPARC (ver. 2.2.0)
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: cryoSPARC (ver. 2.2.0)
Final reconstructionApplied symmetry - Point group: C3 (3 fold cyclic) / Resolution.type: BY AUTHOR / Resolution: 2.61 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: cryoSPARC (ver. 2.2.0) / Number images used: 569723
FSC plot (resolution estimation)

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Atomic model buiding 1

Initial modelPDB ID:

6uyn

RefinementSpace: REAL / Protocol: RIGID BODY FIT / Overall B value: 123.2
Output model

PDB-8diu:
Cryo-EM structure of influenza A virus A/Bayern/7/1995 hemagglutinin bound to CR6261 Fab

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