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- PDB-9rwc: Crystal structure of a sialic acid binding protein, R404A mutant,... -

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Basic information

Entry
Database: PDB / ID: 9rwc
TitleCrystal structure of a sialic acid binding protein, R404A mutant, from Streptococcus pneumoniae bound to Neu5Ac
ComponentsSugar ABC transporter, sugar-binding protein
KeywordsSUGAR BINDING PROTEIN / Sialic acid / Neu5Ac / Periplasmic binding protein / Streptococcus pneumoniae / mutant
Function / homologymaltose binding / maltose transport / maltodextrin transmembrane transport / ATP-binding cassette (ABC) transporter complex, substrate-binding subunit-containing / Bacterial extracellular solute-binding protein / Bacterial extracellular solute-binding protein / Prokaryotic membrane lipoprotein lipid attachment site profile. / N-acetyl-alpha-neuraminic acid / Sugar ABC transporter, sugar-binding protein
Function and homology information
Biological speciesStreptococcus pneumoniae TIGR4 (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 1.797 Å
AuthorsAtkinson, M. / Lukacik, P. / Strain-Damerell, C.M. / Gloster, T.M. / Walsh, M.A.
Funding support United Kingdom, 1items
OrganizationGrant numberCountry
Diamond Light Source United Kingdom
CitationJournal: To Be Published / Year: 2026
Title: Structure of S. pneumoniae sialic acid binding protein; SatA
Authors: Strain-Damerell, C.M. / Atkinson, M. / Meller, C. / Harris, G. / Gloster, T.M. / Lukacik, P. / Walsh, M.A.
History
DepositionJul 9, 2025Deposition site: PDBE / Processing site: PDBE
Revision 1.0Jul 29, 2026Provider: repository / Type: Initial release

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

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Assembly

Deposited unit
A: Sugar ABC transporter, sugar-binding protein
B: Sugar ABC transporter, sugar-binding protein
hetero molecules


Theoretical massNumber of molelcules
Total (without water)97,2626
Polymers96,4522
Non-polymers8114
Water18,2491013
1


  • Idetical with deposited unit
  • defined by author
  • Evidence: gel filtration, peak elution volume corresponds to monomeric mass of chain A or B
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area2730 Å2
ΔGint-17 kcal/mol
Surface area33420 Å2
Unit cell
Length a, b, c (Å)78.255, 61.146, 88.873
Angle α, β, γ (deg.)90.000, 106.127, 90.000
Int Tables number4
Space group name H-MP1211
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails (eV)
11A
21B

NCS domain segments:

Component-ID: 1 / Ens-ID: 1 / Beg auth comp-ID: LYS / Beg label comp-ID: LYS / End auth comp-ID: LYS / End label comp-ID: LYS / Auth seq-ID: 40 - 437 / Label seq-ID: 37 - 434

Dom-IDAuth asym-IDLabel asym-ID
1AA
2BB

NCS ensembles : (Details: Local NCS retraints between domains: 1 2)

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Components

#1: Protein Sugar ABC transporter, sugar-binding protein


Mass: 48225.848 Da / Num. of mol.: 2
Source method: isolated from a genetically manipulated source
Details: R404A mutant
Source: (gene. exp.) Streptococcus pneumoniae TIGR4 (bacteria)
Gene: SP_1683 / Plasmid: pOPINF / Production host: Escherichia coli BL21(DE3) (bacteria) / Strain (production host): BL21(DE3) / Variant (production host): Rosetta / References: UniProt: A0A0H2URD1
#2: Sugar ChemComp-SIA / N-acetyl-alpha-neuraminic acid / N-acetylneuraminic acid / sialic acid / alpha-sialic acid / O-SIALIC ACID


Type: D-saccharide, alpha linking / Mass: 309.270 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: C11H19NO9 / Feature type: SUBJECT OF INVESTIGATION
IdentifierTypeProgram
DNeup5AcaCONDENSED IUPAC CARBOHYDRATE SYMBOLGMML 1.0
N-acetyl-a-D-neuraminic acidCOMMON NAMEGMML 1.0
a-D-Neup5AcIUPAC CARBOHYDRATE SYMBOLPDB-CARE 1.0
Neu5AcSNFG CARBOHYDRATE SYMBOLGMML 1.0
#3: Chemical ChemComp-SO4 / SULFATE ION


Mass: 96.063 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: SO4
#4: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 1013 / Source method: isolated from a natural source / Formula: H2O
Has ligand of interestY
Has protein modificationN

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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: 2.12 Å3/Da / Density % sol: 41.92 %
Crystal growTemperature: 293 K / Method: vapor diffusion, sitting drop / pH: 5.4 / Details: 0.1M NaCit, 3.8M ammonium sulfate, pH 5.4

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Data collection

DiffractionMean temperature: 100 K / Serial crystal experiment: N
Diffraction sourceSource: SYNCHROTRON / Site: Diamond / Beamline: I03 / Wavelength: 0.9762 Å
DetectorType: DECTRIS EIGER2 XE 16M / Detector: PIXEL / Date: Apr 9, 2025
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.9762 Å / Relative weight: 1
ReflectionResolution: 1.797→85.376 Å / Num. obs: 55625 / % possible obs: 92 % / Redundancy: 7.11 % / CC1/2: 0.993 / CC1/2 anomalous: -0.185 / Rmerge(I) obs: 0.1722 / Rpim(I) all: 0.0692 / Rrim(I) all: 0.1858 / AbsDiff over sigma anomalous: 0.692 / Baniso tensor eigenvalue 1: 44.9679 Å2 / Baniso tensor eigenvalue 2: 0 Å2 / Baniso tensor eigenvalue 3: 1.1857 Å2 / Baniso tensor eigenvector 1 ortho1: 0.9959 / Baniso tensor eigenvector 1 ortho2: 0 / Baniso tensor eigenvector 1 ortho3: 0.0905 / Baniso tensor eigenvector 2 ortho1: 0 / Baniso tensor eigenvector 2 ortho2: 1 / Baniso tensor eigenvector 2 ortho3: 0 / Baniso tensor eigenvector 3 ortho1: -0.0905 / Baniso tensor eigenvector 3 ortho2: 0 / Baniso tensor eigenvector 3 ortho3: 0.9959 / Aniso diffraction limit 1: 2.449 Å / Aniso diffraction limit 2: 1.636 Å / Aniso diffraction limit 3: 1.668 Å / Aniso diffraction limit axis 1 ortho1: 0.99338 / Aniso diffraction limit axis 1 ortho2: 0 / Aniso diffraction limit axis 1 ortho3: 0.1148 / Aniso diffraction limit axis 2 ortho1: 0 / Aniso diffraction limit axis 2 ortho2: 1 / Aniso diffraction limit axis 2 ortho3: 0 / Aniso diffraction limit axis 3 ortho1: -0.1148 / Aniso diffraction limit axis 3 ortho2: 0 / Aniso diffraction limit axis 3 ortho3: 0.99338 / Net I/σ(I): 7.78 / Num. measured all: 395309 / Observed signal threshold: 1.2 / Orthogonalization convention: pdb / % possible anomalous: 92.4 / % possible ellipsoidal: 92 / % possible ellipsoidal anomalous: 92.4 / % possible spherical: 73.8 / % possible spherical anomalous: 74.3 / Redundancy anomalous: 3.63 / Signal type: local
Reflection shell
Resolution (Å)Redundancy (%)Rmerge(I) obsMean I/σ(I) obsNum. measured allNum. measured obsNum. unique allNum. unique obsCC1/2CC1/2 anomalousRpim(I) allRrim(I) allAbsDiff over sigma anomalous% possible anomalous% possible ellipsoidal% possible ellipsoidal anomalous% possible spherical% possible spherical anomalousRedundancy anomalous% possible all
5.472-85.3766.810.077314.931893218932278127810.996-0.3980.03170.08370.55910099.810099.81003.6399.8
4.318-5.4717.010.096514.271951319513278227820.994-0.180.03910.10430.61310099.710099.71003.6499.7
3.767-4.3187.340.098914.322040720407278127810.994-0.4060.03910.10640.60810099.610099.61003.7899.6
3.416-3.7677.480.09513.212080520805278127810.995-0.3010.03730.10210.59399.899.599.899.599.83.8499.5
3.168-3.4167.210.111510.992005220052278127810.995-0.2360.04460.12020.64699.499.399.499.399.43.799.3
2.979-3.1686.50.139191807118071278227820.992-0.1140.05890.15130.67899.699.399.699.399.63.3299.3
2.825-2.9796.930.17188.151926219262278127810.989-0.0650.07040.18580.68698.297.398.297.398.23.5397.3
2.693-2.8257.150.20597.411988519885278127810.987-0.1210.08270.2220.69494.192.894.192.894.13.6392.8
2.576-2.6927.280.21817.012025020250278127810.987-0.1070.08670.23490.7088987.58987.5893.6987.5
2.473-2.5767.340.23116.852040920409278227820.984-0.0590.09150.24870.70684.983.584.983.584.93.7183.5
2.38-2.4737.370.25386.432049520495278127810.98-0.1380.10.2730.70781.380.281.378.679.93.7380.2
2.295-2.387.40.26486.332057520575278127810.98-0.1130.1040.28470.71883.582.783.575.376.63.7482.7
2.219-2.2957.350.27616.232042920429278127810.974-0.1130.10910.2970.71286.285.686.272.373.43.7285.6
2.149-2.2196.880.30265.651913419134278227820.972-0.140.1240.32730.72187.586.987.568.569.73.4786.9
2.083-2.1496.860.32955.231908419084278127810.97-0.1620.13490.35630.72690.289.790.265.166.13.4689.7
2.022-2.0837.060.38574.821964319643278127810.948-0.1060.15540.4160.74492.492.192.461.862.73.5792.1
1.963-2.0227.130.43324.411983519835278127810.942-0.0790.17360.4670.75194.293.794.25858.83.693.7
1.909-1.9637.210.51153.842006920069278227820.914-0.0920.20360.55080.73995.394.995.355.255.93.6494.9
1.855-1.9097.250.56853.442015420154278127810.901-0.0580.22560.61210.7549594.99549.249.83.6694.9
1.797-1.8556.580.59313.021830518305278127810.875-0.0480.24350.64230.74387.290.187.241.140.13.4390.1

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Processing

Software
NameVersionClassification
REFMAC5.8.0430refinement
autoPROCdata reduction
autoPROCdata scaling
REFMACphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT / Resolution: 1.797→85.376 Å / Cor.coef. Fo:Fc: 0.919 / Cor.coef. Fo:Fc free: 0.862 / SU B: 3.449 / SU ML: 0.11 / Cross valid method: FREE R-VALUE / ESU R: 0.223 / ESU R Free: 0.191
Details: Hydrogens have been added in their riding positions
RfactorNum. reflection% reflection
Rfree0.2525 2715 4.881 %
Rwork0.1947 52910 -
all0.198 --
obs-55625 73.814 %
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.2 Å / Solvent model: MASK BULK SOLVENT
Displacement parametersBiso mean: 9.143 Å2
Baniso -1Baniso -2Baniso -3
1-0.534 Å2-0 Å2-0.011 Å2
2---0.223 Å20 Å2
3----0.262 Å2
Refinement stepCycle: LAST / Resolution: 1.797→85.376 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms6248 0 52 1013 7313
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0080.0126590
X-RAY DIFFRACTIONr_bond_other_d0.0010.0166117
X-RAY DIFFRACTIONr_angle_refined_deg1.5481.8128957
X-RAY DIFFRACTIONr_angle_other_deg0.5561.77514224
X-RAY DIFFRACTIONr_dihedral_angle_1_deg6.4365840
X-RAY DIFFRACTIONr_dihedral_angle_2_deg7.707510
X-RAY DIFFRACTIONr_dihedral_angle_3_deg12.274101132
X-RAY DIFFRACTIONr_dihedral_angle_6_deg15.13610299
X-RAY DIFFRACTIONr_chiral_restr0.080.2966
X-RAY DIFFRACTIONr_gen_planes_refined0.0080.027727
X-RAY DIFFRACTIONr_gen_planes_other0.0010.021445
X-RAY DIFFRACTIONr_nbd_refined0.2170.21569
X-RAY DIFFRACTIONr_symmetry_nbd_other0.1910.25866
X-RAY DIFFRACTIONr_nbtor_refined0.1890.23380
X-RAY DIFFRACTIONr_symmetry_nbtor_other0.0760.23243
X-RAY DIFFRACTIONr_xyhbond_nbd_refined0.2050.2869
X-RAY DIFFRACTIONr_symmetry_xyhbond_nbd_other0.0160.21
X-RAY DIFFRACTIONr_symmetry_nbd_refined0.1940.227
X-RAY DIFFRACTIONr_nbd_other0.190.2101
X-RAY DIFFRACTIONr_symmetry_xyhbond_nbd_refined0.1810.277
X-RAY DIFFRACTIONr_mcbond_it0.8330.8923270
X-RAY DIFFRACTIONr_mcbond_other0.8310.8923270
X-RAY DIFFRACTIONr_mcangle_it1.3631.5944107
X-RAY DIFFRACTIONr_mcangle_other1.3631.5964108
X-RAY DIFFRACTIONr_scbond_it1.4161.0293320
X-RAY DIFFRACTIONr_scbond_other1.4121.0233311
X-RAY DIFFRACTIONr_scangle_it2.3181.8084839
X-RAY DIFFRACTIONr_scangle_other2.3161.7964828
X-RAY DIFFRACTIONr_lrange_it4.31211.1298428
X-RAY DIFFRACTIONr_lrange_other3.9629.8978089
X-RAY DIFFRACTIONr_ncsr_local_group_10.0630.0513827
Refine LS restraints NCS
Ens-IDDom-IDAuth asym-IDRefine-IDTypeRms dev position (Å)Weight position
11AX-RAY DIFFRACTIONLocal ncs0.062740.05009
12BX-RAY DIFFRACTIONLocal ncs0.062740.05009
LS refinement shell

Refine-ID: X-RAY DIFFRACTION / Total num. of bins used: 20

Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRfactor allNum. reflection allFsc freeFsc work% reflection obs (%)WRfactor Rwork
1.797-1.8440.2321080.21920970.21955530.9640.97339.70830.198
1.844-1.8940.2651290.21824870.22154120.9540.9748.3370.198
1.894-1.9490.2531350.18926290.19251930.9560.97553.22550.164
1.949-2.0090.2491300.1828220.18351360.9580.97757.47660.155
2.009-2.0750.2511450.18728610.1949330.9580.97660.93660.163
2.075-2.1480.2671430.17829720.18247670.9540.97865.34510.152
2.148-2.2290.2441450.18130470.18346460.960.97768.70430.154
2.229-2.320.2371570.1730830.17344430.9650.9872.92370.147
2.32-2.4230.2311630.17731080.1842750.9620.97976.51460.153
2.423-2.5410.2411870.17631310.1840990.9610.97880.94660.152
2.541-2.6780.2851540.20632320.2138900.9520.9787.04370.182
2.678-2.8410.2251680.18332370.18536790.9650.97792.55230.159
2.841-3.0370.251720.19732170.234600.9580.97497.9480.178
3.037-3.2790.2791590.19230590.19632450.9470.97799.16790.182
3.279-3.5920.2171580.18627980.18729810.9670.9899.16140.18
3.592-4.0150.2561330.18625460.18926900.960.97899.59110.183
4.015-4.6340.2431170.20122700.20323960.9630.97699.62440.202
4.634-5.6720.259960.22719300.22920350.9610.97499.55770.234
5.672-8.0040.327690.26415150.26615880.9440.96599.74810.279
8.004-85.3760.352470.2818680.2859170.930.95499.78190.341

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