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- PDB-4b7r: H1N1 2009 Pandemic Influenza Virus: Resistance of the I223R Neura... -

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Basic information

Entry
Database: PDB / ID: 4b7r
TitleH1N1 2009 Pandemic Influenza Virus: Resistance of the I223R Neuraminidase Mutant Explained by Kinetic and Structural Analysis
ComponentsNEURAMINIDASE
KeywordsHYDROLASE / NEURAMINIDASE INHIBITOR / NAI / NAIS / ZANAMIVIR / RESISTANCE / ANTIVIRAL RESISTANCE
Function / homology
Function and homology information


exo-alpha-(2->3)-sialidase activity / exo-alpha-(2->6)-sialidase activity / exo-alpha-(2->8)-sialidase activity / exo-alpha-sialidase / viral budding from plasma membrane / carbohydrate metabolic process / host cell plasma membrane / virion membrane / membrane / metal ion binding
Similarity search - Function
Sialidase, Influenza viruses A/B / Glycoside hydrolase, family 34 / Neuraminidase / Neuraminidase - #10 / Sialidase superfamily / 6 Propeller / Neuraminidase / Mainly Beta
Similarity search - Domain/homology
Chem-G39 / Neuraminidase
Similarity search - Component
Biological speciesINFLUENZA A VIRUS
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 1.9 Å
AuthorsLiu, J. / van der Vries, E. / Vachieri, S.G. / Xiong, X. / Collins, P.J. / Walker, P.A. / Haire, L.F. / Hay, A.J. / Schutten, M. / Osterhaus, A.D.M.E. ...Liu, J. / van der Vries, E. / Vachieri, S.G. / Xiong, X. / Collins, P.J. / Walker, P.A. / Haire, L.F. / Hay, A.J. / Schutten, M. / Osterhaus, A.D.M.E. / Martin, S.R. / Boucher, C.A.B. / Skehel, J.J. / Gamblin, S.J.
CitationJournal: Plos Pathog. / Year: 2012
Title: H1N1 2009 Pandemic Influenza Virus: Resistance of the I223R Neuraminidase Mutant Explained by Kinetic and Structural Analysis
Authors: Van Der Vries, E. / Collins, P.J. / Vachieri, S.G. / Xiong, X. / Liu, J. / Walker, P.A. / Haire, L.F. / Hay, A.J. / Schutten, M. / Osterhaus, A.D.M.E. / Martin, S.R. / Boucher, C.A.B. / ...Authors: Van Der Vries, E. / Collins, P.J. / Vachieri, S.G. / Xiong, X. / Liu, J. / Walker, P.A. / Haire, L.F. / Hay, A.J. / Schutten, M. / Osterhaus, A.D.M.E. / Martin, S.R. / Boucher, C.A.B. / Skehel, J.J. / Gamblin, S.J.
History
DepositionAug 21, 2012Deposition site: PDBE / Processing site: PDBE
Revision 1.0Oct 3, 2012Provider: repository / Type: Initial release
Revision 1.1Oct 10, 2012Group: Database references
Revision 1.2Jul 17, 2013Group: Atomic model / Derived calculations / Non-polymer description
Revision 2.0Jul 29, 2020Group: Advisory / Atomic model ...Advisory / Atomic model / Data collection / Derived calculations / Other / Structure summary
Category: atom_site / chem_comp ...atom_site / chem_comp / entity / pdbx_branch_scheme / pdbx_chem_comp_identifier / pdbx_database_status / pdbx_entity_branch / pdbx_entity_branch_descriptor / pdbx_entity_branch_link / pdbx_entity_branch_list / pdbx_entity_nonpoly / pdbx_nonpoly_scheme / pdbx_struct_assembly_gen / pdbx_struct_conn_angle / pdbx_unobs_or_zero_occ_atoms / struct_asym / struct_conn / struct_site / struct_site_gen
Item: _atom_site.B_iso_or_equiv / _atom_site.Cartn_x ..._atom_site.B_iso_or_equiv / _atom_site.Cartn_x / _atom_site.Cartn_y / _atom_site.Cartn_z / _atom_site.auth_asym_id / _atom_site.auth_atom_id / _atom_site.auth_comp_id / _atom_site.auth_seq_id / _atom_site.label_asym_id / _atom_site.label_atom_id / _atom_site.label_comp_id / _atom_site.label_entity_id / _atom_site.type_symbol / _chem_comp.name / _chem_comp.type / _pdbx_database_status.status_code_sf / _pdbx_entity_nonpoly.entity_id / _pdbx_entity_nonpoly.name / _pdbx_struct_assembly_gen.asym_id_list / _pdbx_struct_conn_angle.ptnr1_auth_comp_id / _pdbx_struct_conn_angle.ptnr1_auth_seq_id / _pdbx_struct_conn_angle.ptnr1_label_asym_id / _pdbx_struct_conn_angle.ptnr1_label_atom_id / _pdbx_struct_conn_angle.ptnr1_label_comp_id / _pdbx_struct_conn_angle.ptnr1_label_seq_id / _pdbx_struct_conn_angle.ptnr2_label_asym_id / _pdbx_struct_conn_angle.ptnr3_auth_comp_id / _pdbx_struct_conn_angle.ptnr3_auth_seq_id / _pdbx_struct_conn_angle.ptnr3_label_asym_id / _pdbx_struct_conn_angle.ptnr3_label_atom_id / _pdbx_struct_conn_angle.ptnr3_label_comp_id / _pdbx_struct_conn_angle.ptnr3_label_seq_id / _pdbx_struct_conn_angle.value / _pdbx_unobs_or_zero_occ_atoms.label_asym_id / _struct_conn.conn_type_id / _struct_conn.id / _struct_conn.pdbx_dist_value / _struct_conn.pdbx_leaving_atom_flag / _struct_conn.pdbx_role / _struct_conn.ptnr1_auth_asym_id / _struct_conn.ptnr1_auth_comp_id / _struct_conn.ptnr1_auth_seq_id / _struct_conn.ptnr1_label_asym_id / _struct_conn.ptnr1_label_atom_id / _struct_conn.ptnr1_label_comp_id / _struct_conn.ptnr1_label_seq_id / _struct_conn.ptnr2_auth_asym_id / _struct_conn.ptnr2_auth_comp_id / _struct_conn.ptnr2_auth_seq_id / _struct_conn.ptnr2_label_asym_id / _struct_conn.ptnr2_label_atom_id / _struct_conn.ptnr2_label_comp_id / _struct_conn.ptnr2_label_seq_id
Description: Carbohydrate remediation / Provider: repository / Type: Remediation

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: NEURAMINIDASE
B: NEURAMINIDASE
C: NEURAMINIDASE
D: NEURAMINIDASE
hetero molecules


Theoretical massNumber of molelcules
Total (without water)176,05233
Polymers170,5624
Non-polymers5,49029
Water17,222956
1


  • Idetical with deposited unit
  • defined by software
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area20530 Å2
ΔGint-106.1 kcal/mol
Surface area44810 Å2
MethodPISA
Unit cell
Length a, b, c (Å)82.650, 148.820, 166.600
Angle α, β, γ (deg.)90.00, 90.00, 90.00
Int Tables number19
Space group name H-MP212121

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Components

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Protein , 1 types, 4 molecules ABCD

#1: Protein
NEURAMINIDASE /


Mass: 42640.551 Da / Num. of mol.: 4 / Source method: isolated from a natural source / Details: OSELTAMIVIR LIGAND BOUND
Source: (natural) INFLUENZA A VIRUS (A/CALIFORNIA/07/2009(H1N1))
References: UniProt: C7FH46

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Sugars , 2 types, 13 molecules

#2: Polysaccharide 2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose


Type: oligosaccharide / Mass: 424.401 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
DescriptorTypeProgram
DGlcpNAcb1-4DGlcpNAcb1-Glycam Condensed SequenceGMML 1.0
WURCS=2.0/1,2,1/[a2122h-1b_1-5_2*NCC/3=O]/1-1/a4-b1WURCSPDB2Glycan 1.1.0
[]{[(4+1)][b-D-GlcpNAc]{[(4+1)][b-D-GlcpNAc]{}}}LINUCSPDB-CARE
#6: Sugar
ChemComp-NAG / 2-acetamido-2-deoxy-beta-D-glucopyranose / N-Acetylglucosamine


Type: D-saccharide, beta linking / Mass: 221.208 Da / Num. of mol.: 12
Source method: isolated from a genetically manipulated source
Formula: C8H15NO6
IdentifierTypeProgram
DGlcpNAcbCONDENSED IUPAC CARBOHYDRATE SYMBOLGMML 1.0
N-acetyl-b-D-glucopyranosamineCOMMON NAMEGMML 1.0
b-D-GlcpNAcIUPAC CARBOHYDRATE SYMBOLPDB-CARE 1.0
GlcNAcSNFG CARBOHYDRATE SYMBOLGMML 1.0

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Non-polymers , 4 types, 972 molecules

#3: Chemical
ChemComp-CA / CALCIUM ION


Mass: 40.078 Da / Num. of mol.: 8 / Source method: obtained synthetically / Formula: Ca
#4: Chemical
ChemComp-G39 / (3R,4R,5S)-4-(acetylamino)-5-amino-3-(pentan-3-yloxy)cyclohex-1-ene-1-carboxylic acid / Oseltamivir carboxylate / Oseltamivir


Mass: 284.351 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: C14H24N2O4 / Comment: medication, antivirus*YM
#5: Chemical
ChemComp-EPE / 4-(2-HYDROXYETHYL)-1-PIPERAZINE ETHANESULFONIC ACID / HEPES / HEPES


Mass: 238.305 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: C8H18N2O4S / Comment: pH buffer*YM
#7: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 956 / Source method: isolated from a natural source / Formula: H2O

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Details

Sequence detailsRESIDUES 1-82 WERE PROTEOLYTICALY CLEAVED USING BROMELAIN DURING PURIFICATION

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Experimental details

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Experiment

ExperimentMethod: X-RAY DIFFRACTION / Number of used crystals: 1

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Sample preparation

CrystalDensity Matthews: 3.05 Å3/Da / Density % sol: 59.66 % / Description: NONE
Crystal growDetails: 15% PEG3350, 0.1M BIS-TRIS PROPANE AND 0.1M SODIUM ACETATE BUFFER (PH 4.6)

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Data collection

DiffractionMean temperature: 77 K
Diffraction sourceSource: SYNCHROTRON / Site: Diamond / Beamline: I02 / Wavelength: 0.9795
DetectorType: ADSC QUANTUM 315 / Detector: CCD / Date: Feb 24, 2010
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.9795 Å / Relative weight: 1
ReflectionResolution: 1.9→30 Å / Num. obs: 163254 / % possible obs: 99.7 % / Observed criterion σ(I): 2 / Redundancy: 6.4 % / Biso Wilson estimate: 28.07 Å2 / Rmerge(I) obs: 0.12 / Net I/σ(I): 13.5
Reflection shellResolution: 1.9→1.99 Å / Redundancy: 4.8 % / Rmerge(I) obs: 0.73 / Mean I/σ(I) obs: 3 / % possible all: 98

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Processing

Software
NameVersionClassification
PHENIX(PHENIX.REFINE)refinement
DENZOdata reduction
SCALEPACKdata scaling
PHASERphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT / Resolution: 1.9→29.818 Å / SU ML: 0.17 / σ(F): 100 / Phase error: 17.4 / Stereochemistry target values: ML
RfactorNum. reflection% reflection
Rfree0.1732 8184 5 %
Rwork0.1457 --
obs0.1471 163045 99.66 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL / Bsol: 34.056 Å2 / ksol: 0.353 e/Å3
Displacement parametersBiso mean: 28.6 Å2
Refinement stepCycle: LAST / Resolution: 1.9→29.818 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms11976 0 344 956 13276
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.00712685
X-RAY DIFFRACTIONf_angle_d1.1917201
X-RAY DIFFRACTIONf_dihedral_angle_d14.7794571
X-RAY DIFFRACTIONf_chiral_restr0.0871817
X-RAY DIFFRACTIONf_plane_restr0.0052206
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
1.9-1.92160.30452290.30054844X-RAY DIFFRACTION94
1.9216-1.94420.26472590.22595024X-RAY DIFFRACTION98
1.9442-1.96790.25042680.20385041X-RAY DIFFRACTION99
1.9679-1.99280.19792450.1835180X-RAY DIFFRACTION100
1.9928-2.0190.22382610.17565112X-RAY DIFFRACTION100
2.019-2.04670.20762670.17315130X-RAY DIFFRACTION100
2.0467-2.07590.2252770.18055142X-RAY DIFFRACTION100
2.0759-2.10690.21152360.16215179X-RAY DIFFRACTION100
2.1069-2.13980.1952700.14825141X-RAY DIFFRACTION100
2.1398-2.17490.17192510.14895133X-RAY DIFFRACTION100
2.1749-2.21240.18622800.15575139X-RAY DIFFRACTION100
2.2124-2.25260.2273020.19245143X-RAY DIFFRACTION100
2.2526-2.29590.19792710.16315099X-RAY DIFFRACTION100
2.2959-2.34270.19382800.14955156X-RAY DIFFRACTION100
2.3427-2.39360.18572830.14535129X-RAY DIFFRACTION100
2.3936-2.44930.19163040.14595146X-RAY DIFFRACTION100
2.4493-2.51050.17922890.1475141X-RAY DIFFRACTION100
2.5105-2.57840.17933070.14595103X-RAY DIFFRACTION100
2.5784-2.65420.1922710.14745158X-RAY DIFFRACTION100
2.6542-2.73980.1762410.15115204X-RAY DIFFRACTION100
2.7398-2.83770.17772760.14555183X-RAY DIFFRACTION100
2.8377-2.95120.19582900.14575181X-RAY DIFFRACTION100
2.9512-3.08540.16182740.1385175X-RAY DIFFRACTION100
3.0854-3.24780.17282730.14055192X-RAY DIFFRACTION100
3.2478-3.4510.15712660.13485236X-RAY DIFFRACTION100
3.451-3.7170.15012680.135232X-RAY DIFFRACTION100
3.717-4.09030.13932910.12195204X-RAY DIFFRACTION100
4.0903-4.68020.13142530.11325318X-RAY DIFFRACTION100
4.6802-5.88910.15293080.13245296X-RAY DIFFRACTION100
5.8891-29.82210.17582940.16325500X-RAY DIFFRACTION100
Refinement TLS params.

Method: refined / Refine-ID: X-RAY DIFFRACTION

IDL112)L122)L132)L222)L232)L332)S11 (Å °)S12 (Å °)S13 (Å °)S21 (Å °)S22 (Å °)S23 (Å °)S31 (Å °)S32 (Å °)S33 (Å °)T112)T122)T132)T222)T232)T332)Origin x (Å)Origin y (Å)Origin z (Å)
10.48630.02360.2670.6021-0.09830.5109-0.0241-0.0050.07910.0593-0.0192-0.0454-0.1150.02620.06040.2237-0.00320.01190.2080.02180.232820.165412.3301-29.5125
20.7447-0.08880.32220.4948-0.07190.8443-0.07970.01270.20130.0636-0.0303-0.0698-0.2990.05550.110.30120.0014-0.02240.18280.03490.262216.520627.5103-34.2489
30.74990.15210.0971.01150.05371.133-0.02990.0050.29240.1033-0.0561-0.3209-0.40.27460.06960.3623-0.09-0.07520.27090.04530.363433.993328.377-24.2802
40.62470.03470.60.6053-0.19010.6625-0.08370.00460.11560.08940.0402-0.1267-0.13540.16740.05260.237-0.0238-0.0080.19860.03390.241727.531214.8155-25.2962
50.2995-0.020.08620.604-0.00010.4413-0.03110.04370.05210.0171-0.00170.0269-0.0641-0.01350.02450.20030.01-0.01050.21980.00150.212423.334-3.1573-11.5442
60.46440.05530.23860.4560.02880.8541-0.0279-0.00160.04310.1009-0.0121-0.0714-0.02310.0730.03870.18630.0111-0.00920.1801-0.00660.178430.7506-4.51941.8146
70.54120.31520.33830.4854-0.06780.4449-0.0058-0.04320.04170.0347-0.00570.0364-0.0049-0.04690.00560.1840.01720.02420.1888-0.01670.18656.5886-20.4943-21.4465
80.49910.01650.02750.16810.10670.3699-0.06870.05250.07620.0566-0.03540.0711-0.029-0.14640.09070.2160.01040.01310.2145-0.01030.23027.7836-15.3185-15.3532
90.5940.19320.01230.58280.10510.49780.0589-0.0935-0.01660.1528-0.03340.05920.0958-0.1035-0.03070.2448-0.00970.01780.2098-0.00030.18524.5403-28.7159-3.6097
100.61670.27180.40180.6595-0.14030.65630.063-0.0476-0.07710.0866-0.02190.08080.1699-0.1048-0.04070.2133-0.02680.02080.1715-0.00120.1815-5.6377-38.0229-17.0224
110.73880.4273-0.09040.7903-0.32680.9203-0.07890.2133-0.183-0.07060.08240.06370.1593-0.18170.01410.2413-0.02020.0140.2388-0.04310.2401-7.5842-37.0855-27.6917
120.57960.28670.36220.7389-0.38041.06760.03470.09520.0066-0.0054-0.01260.0060.0508-0.0636-0.03010.1874-0.00670.00950.169-0.01530.17471.472-24.3875-27.4426
130.31110.1860.05650.5806-0.28670.38850.0005-0.00950.0121-0.0044-0.0020.0352-0.0396-0.05330.00390.21240.02850.01270.23620.00280.19012.0909-2.8327-39.7167
140.3474-0.0189-0.08160.5168-0.38320.46840.0153-0.01020.10310.0330.02310.0471-0.0309-0.0533-0.06130.21340.02010.01860.2337-0.00050.2202-5.662-9.3997-33.1433
150.8322-0.3466-0.50241.56410.3311.06810.05240.00270.08780.01860.0170.2311-0.0629-0.3186-0.04970.17450.01310.00410.27130.02610.2373-15.3422-8.907-41.9254
160.47030.03790.00390.6633-0.2440.59110.0043-0.0310.04710.00460.07330.2234-0.0674-0.2782-0.06810.19990.01220.00630.32850.03680.2886-19.1907-4.4719-47.1605
170.80830.3729-0.13320.7886-0.52110.7103-0.03420.33380.0478-0.21880.02930.11170.0055-0.2128-0.02760.22530.0301-0.02360.35530.03750.2516-12.60820.733-60.3692
180.98460.0122-0.09960.67680.14120.5111-0.01620.32170.1629-0.13870.00860.083-0.1301-0.13750.01740.21320.04730.00190.250.05310.2088-5.64628.6073-56.5159
190.73310.29560.33250.8279-0.29880.59010.04490.06020.1126-0.0995-0.0166-0.0078-0.0728-0.0031-0.03560.19890.02010.01560.21250.02370.17473.52733.5855-45.6374
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection details
1X-RAY DIFFRACTION1(CHAIN A AND RESID 83:156)
2X-RAY DIFFRACTION2(CHAIN A AND RESID 157:349)
3X-RAY DIFFRACTION3(CHAIN A AND RESID 350:402)
4X-RAY DIFFRACTION4(CHAIN A AND RESID 403:469)
5X-RAY DIFFRACTION5(CHAIN B AND RESID 83:189)
6X-RAY DIFFRACTION6(CHAIN B AND RESID 190:469)
7X-RAY DIFFRACTION7(CHAIN C AND RESID 83:156)
8X-RAY DIFFRACTION8(CHAIN C AND RESID 157:189)
9X-RAY DIFFRACTION9(CHAIN C AND RESID 190:279)
10X-RAY DIFFRACTION10(CHAIN C AND RESID 280:376)
11X-RAY DIFFRACTION11(CHAIN C AND RESID 377:402)
12X-RAY DIFFRACTION12(CHAIN C AND RESID 403:469)
13X-RAY DIFFRACTION13(CHAIN D AND RESID 83:156)
14X-RAY DIFFRACTION14(CHAIN D AND RESID 157:217)
15X-RAY DIFFRACTION15(CHAIN D AND RESID 218:245)
16X-RAY DIFFRACTION16(CHAIN D AND RESID 246:311)
17X-RAY DIFFRACTION17(CHAIN D AND RESID 312:349)
18X-RAY DIFFRACTION18(CHAIN D AND RESID 350:402)
19X-RAY DIFFRACTION19(CHAIN D AND RESID 403:469)

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