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- PDB-8p1s: Bifidobacterium asteroides alpha-L-fucosidase (TT1819) in complex... -

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Basic information

Entry
Database: PDB / ID: 8p1s
TitleBifidobacterium asteroides alpha-L-fucosidase (TT1819) in complex with fucose.
ComponentsBifidobacterium asteroides alpha-L-fucosidase (TT1819)
KeywordsSUGAR BINDING PROTEIN / glycosidase hydrolase / fucosidase / GH29
Function / homologyalpha-L-fucopyranose / IMIDAZOLE
Function and homology information
Biological speciesBifidobacterium asteroides (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 1.9 Å
AuthorsOwen, C.D. / Penner, M. / Gascuena, A.N. / Wu, H. / Hernando, P.J. / Monaco, S. / Le Gall, G. / Gardner, R. / Ndeh, D. / Urbanowicz, P.A. ...Owen, C.D. / Penner, M. / Gascuena, A.N. / Wu, H. / Hernando, P.J. / Monaco, S. / Le Gall, G. / Gardner, R. / Ndeh, D. / Urbanowicz, P.A. / Spencer, D.I.R. / Walsh, M.A. / Angulo, J. / Juge, N.
Funding support United Kingdom, Spain, European Union, 4items
OrganizationGrant numberCountry
Biotechnology and Biological Sciences Research Council (BBSRC)BB/P010660/1 United Kingdom
Spanish Ministry of Science, Innovation, and UniversitiesPID2019-109395GB-I00 Spain
H2020 Marie Curie Actions of the European Commission814102European Union
UK Research and Innovation (UKRI)BB/M029042/ United Kingdom
CitationJournal: To Be Published
Title: Exploring sequence, structure and function of microbial fucosidases from glycoside hydrolase GH29 family
Authors: Owen, C.D. / Penner, M. / Gascuena, A.N. / Wu, H. / Hernando, P.J. / Monaco, S. / Le Gall, G. / Gardner, R. / Ndeh, D. / Urbanowicz, P.A. / Spencer, D.I.R. / Walsh, M.A. / Angulo, J. / Juge, N.
History
DepositionMay 12, 2023Deposition site: PDBE / Processing site: PDBE
Revision 1.0May 22, 2024Provider: repository / Type: Initial release

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Bifidobacterium asteroides alpha-L-fucosidase (TT1819)
B: Bifidobacterium asteroides alpha-L-fucosidase (TT1819)
C: Bifidobacterium asteroides alpha-L-fucosidase (TT1819)
D: Bifidobacterium asteroides alpha-L-fucosidase (TT1819)
E: Bifidobacterium asteroides alpha-L-fucosidase (TT1819)
F: Bifidobacterium asteroides alpha-L-fucosidase (TT1819)
hetero molecules


Theoretical massNumber of molelcules
Total (without water)234,97222
Polymers233,2026
Non-polymers1,77016
Water13,205733
1


  • Idetical with deposited unit
  • defined by author
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area16690 Å2
ΔGint21 kcal/mol
Surface area67200 Å2
Unit cell
Length a, b, c (Å)89.058, 142.765, 167.202
Angle α, β, γ (deg.)90.000, 90.000, 90.000
Int Tables number19
Space group name H-MP212121
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11A
21A
32A
42A
53A
63A
74A
84A
95A
105A
116A
126A
137A
147A
158A
168A
179A
189A
1910A
2010A
2111A
2211A
2312A
2412A
2513A
2613A
2714A
2814A
2915A
3015A

NCS domain segments:

Beg auth comp-ID: SER / Beg label comp-ID: SER / End auth comp-ID: ALA / End label comp-ID: ALA / Auth asym-ID: A / Label asym-ID: A / Auth seq-ID: 22 - 362 / Label seq-ID: 2 - 342

Dom-IDComponent-IDEns-ID
111
211
322
422
533
633
744
844
955
1055
1166
1266
1377
1477
1588
1688
1799
1899
191010
201010
211111
221111
231212
241212
251313
261313
271414
281414
291515
301515

NCS ensembles :
IDDetails
1Local NCS retraints between domains: 1 2
2Local NCS retraints between domains: 3 4
3Local NCS retraints between domains: 5 6
4Local NCS retraints between domains: 7 8
5Local NCS retraints between domains: 9 10
6Local NCS retraints between domains: 11 12
7Local NCS retraints between domains: 13 14
8Local NCS retraints between domains: 15 16
9Local NCS retraints between domains: 17 18
10Local NCS retraints between domains: 19 20
11Local NCS retraints between domains: 21 22
12Local NCS retraints between domains: 23 24
13Local NCS retraints between domains: 25 26
14Local NCS retraints between domains: 27 28
15Local NCS retraints between domains: 29 30

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Components

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Protein / Sugars , 2 types, 12 molecules ABCDEF

#1: Protein
Bifidobacterium asteroides alpha-L-fucosidase (TT1819)


Mass: 38867.016 Da / Num. of mol.: 6
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Bifidobacterium asteroides (bacteria) / Production host: Escherichia coli (E. coli)
#3: Sugar
ChemComp-FUC / alpha-L-fucopyranose / alpha-L-fucose / 6-deoxy-alpha-L-galactopyranose / L-fucose / fucose / Fucose


Type: L-saccharide, alpha linking / Mass: 164.156 Da / Num. of mol.: 6 / Source method: obtained synthetically / Formula: C6H12O5 / Feature type: SUBJECT OF INVESTIGATION
IdentifierTypeProgram
LFucpaCONDENSED IUPAC CARBOHYDRATE SYMBOLGMML 1.0
a-L-fucopyranoseCOMMON NAMEGMML 1.0
a-L-FucpIUPAC CARBOHYDRATE SYMBOLPDB-CARE 1.0
FucSNFG CARBOHYDRATE SYMBOLGMML 1.0

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Non-polymers , 5 types, 743 molecules

#2: Chemical
ChemComp-EDO / 1,2-ETHANEDIOL / ETHYLENE GLYCOL / Ethylene glycol


Mass: 62.068 Da / Num. of mol.: 5 / Source method: obtained synthetically / Formula: C2H6O2
#4: Chemical ChemComp-TRS / 2-AMINO-2-HYDROXYMETHYL-PROPANE-1,3-DIOL / TRIS BUFFER / Tris


Mass: 122.143 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: C4H12NO3 / Comment: pH buffer*YM
#5: Chemical ChemComp-IMD / IMIDAZOLE / Imidazole


Mass: 69.085 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: C3H5N2
#6: Chemical ChemComp-GOL / GLYCEROL / GLYCERIN / PROPANE-1,2,3-TRIOL / Glycerol


Mass: 92.094 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C3H8O3
#7: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 733 / Source method: isolated from a natural source / Formula: H2O

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Details

Has ligand of interestY

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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: 2.32 Å3/Da / Density % sol: 46.97 %
Crystal growTemperature: 293 K / Method: vapor diffusion, sitting drop
Details: 0.12M Diethylene glycol; 0.12M Triethylene glycol; 0.12M Tetraethylene glycol; 0.12M Pentaethylene glycol, 0.1M Tris (base); BICINE pH 8.5, 10% v/v MPD; 10% PEG 1000; 10% w/v PEG 3350

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Data collection

DiffractionMean temperature: 100 K / Serial crystal experiment: N
Diffraction sourceSource: SYNCHROTRON / Site: Diamond / Beamline: I24 / Wavelength: 0.97625 Å
DetectorType: DECTRIS PILATUS3 X 6M / Detector: PIXEL / Date: Jul 5, 2017
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.97625 Å / Relative weight: 1
ReflectionResolution: 1.9→108.56 Å / Num. obs: 167861 / % possible obs: 100 % / Redundancy: 17.5 % / CC1/2: 0.998 / Rmerge(I) obs: 0.246 / Rpim(I) all: 0.086 / Rrim(I) all: 0.262 / Net I/σ(I): 9.2
Reflection shell

Diffraction-ID: 1

Resolution (Å)Redundancy (%)Rmerge(I) obsNum. unique obsCC1/2Rpim(I) allRrim(I) all
10.41-108.5615.50.0511678610.9990.0180.054
1.9-1.9314.99.94682350.2153.86610.726

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Processing

Software
NameVersionClassification
REFMAC5.8.0405refinement
REFMAC5.8.0405refinement
xia2data reduction
DIALSdata scaling
PHASERphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT / Resolution: 1.9→83.741 Å / Cor.coef. Fo:Fc: 0.972 / Cor.coef. Fo:Fc free: 0.96 / SU B: 9.396 / SU ML: 0.123 / Cross valid method: FREE R-VALUE / ESU R: 0.142 / ESU R Free: 0.131
Details: Hydrogens have been added in their riding positions
RfactorNum. reflection% reflection
Rfree0.2069 8366 4.988 %
Rwork0.1689 159354 -
all0.171 --
obs-167720 99.95 %
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: 38.507 Å2
Baniso -1Baniso -2Baniso -3
1--1.975 Å2-0 Å20 Å2
2--1.839 Å2-0 Å2
3---0.136 Å2
Refinement stepCycle: LAST / Resolution: 1.9→83.741 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms16170 0 118 733 17021
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0140.01216838
X-RAY DIFFRACTIONr_bond_other_d0.0010.01615015
X-RAY DIFFRACTIONr_angle_refined_deg1.9241.64822849
X-RAY DIFFRACTIONr_angle_other_deg0.6461.57834629
X-RAY DIFFRACTIONr_dihedral_angle_1_deg6.66852024
X-RAY DIFFRACTIONr_dihedral_angle_2_deg14.914577
X-RAY DIFFRACTIONr_dihedral_angle_3_deg15.236102703
X-RAY DIFFRACTIONr_dihedral_angle_6_deg16.69710872
X-RAY DIFFRACTIONr_chiral_restr0.0980.22344
X-RAY DIFFRACTIONr_gen_planes_refined0.0110.0219883
X-RAY DIFFRACTIONr_gen_planes_other0.0010.024061
X-RAY DIFFRACTIONr_nbd_refined0.2290.23177
X-RAY DIFFRACTIONr_symmetry_nbd_other0.1940.214296
X-RAY DIFFRACTIONr_nbtor_refined0.1880.28427
X-RAY DIFFRACTIONr_symmetry_nbtor_other0.0830.28621
X-RAY DIFFRACTIONr_xyhbond_nbd_refined0.150.2843
X-RAY DIFFRACTIONr_symmetry_xyhbond_nbd_other0.0610.22
X-RAY DIFFRACTIONr_symmetry_nbd_refined0.2380.212
X-RAY DIFFRACTIONr_nbd_other0.3080.253
X-RAY DIFFRACTIONr_symmetry_xyhbond_nbd_refined0.2280.26
X-RAY DIFFRACTIONr_mcbond_it3.2822.5858095
X-RAY DIFFRACTIONr_mcbond_other3.2732.5868095
X-RAY DIFFRACTIONr_mcangle_it4.1924.62110118
X-RAY DIFFRACTIONr_mcangle_other4.1924.62210119
X-RAY DIFFRACTIONr_scbond_it4.5952.9028743
X-RAY DIFFRACTIONr_scbond_other4.5952.9028744
X-RAY DIFFRACTIONr_scangle_it6.3025.17512731
X-RAY DIFFRACTIONr_scangle_other6.3025.17512732
X-RAY DIFFRACTIONr_lrange_it7.31325.57719434
X-RAY DIFFRACTIONr_lrange_other7.30925.53719338
X-RAY DIFFRACTIONr_ncsr_local_group_10.0660.0511826
X-RAY DIFFRACTIONr_ncsr_local_group_20.0630.0511854
X-RAY DIFFRACTIONr_ncsr_local_group_30.0670.0511778
X-RAY DIFFRACTIONr_ncsr_local_group_40.0670.0511801
X-RAY DIFFRACTIONr_ncsr_local_group_50.0740.0511718
X-RAY DIFFRACTIONr_ncsr_local_group_60.0650.0511787
X-RAY DIFFRACTIONr_ncsr_local_group_70.0660.0511786
X-RAY DIFFRACTIONr_ncsr_local_group_80.060.0511855
X-RAY DIFFRACTIONr_ncsr_local_group_90.070.0511746
X-RAY DIFFRACTIONr_ncsr_local_group_100.060.0511830
X-RAY DIFFRACTIONr_ncsr_local_group_110.0530.0511888
X-RAY DIFFRACTIONr_ncsr_local_group_120.0580.0511818
X-RAY DIFFRACTIONr_ncsr_local_group_130.0550.0512002
X-RAY DIFFRACTIONr_ncsr_local_group_140.0570.0511981
X-RAY DIFFRACTIONr_ncsr_local_group_150.0580.0511787
Refine LS restraints NCS
Ens-IDDom-IDAuth asym-IDRefine-IDTypeRms dev position (Å)Weight position
11AX-RAY DIFFRACTIONLocal ncs0.066160.0501
12AX-RAY DIFFRACTIONLocal ncs0.066160.0501
23AX-RAY DIFFRACTIONLocal ncs0.062840.05009
24AX-RAY DIFFRACTIONLocal ncs0.062840.05009
35AX-RAY DIFFRACTIONLocal ncs0.066520.05009
36AX-RAY DIFFRACTIONLocal ncs0.066520.05009
47AX-RAY DIFFRACTIONLocal ncs0.06720.05009
48AX-RAY DIFFRACTIONLocal ncs0.06720.05009
59AX-RAY DIFFRACTIONLocal ncs0.073740.05009
510AX-RAY DIFFRACTIONLocal ncs0.073740.05009
611AX-RAY DIFFRACTIONLocal ncs0.065080.0501
612AX-RAY DIFFRACTIONLocal ncs0.065080.0501
713AX-RAY DIFFRACTIONLocal ncs0.065770.0501
714AX-RAY DIFFRACTIONLocal ncs0.065770.0501
815AX-RAY DIFFRACTIONLocal ncs0.05990.0501
816AX-RAY DIFFRACTIONLocal ncs0.05990.0501
917AX-RAY DIFFRACTIONLocal ncs0.069580.0501
918AX-RAY DIFFRACTIONLocal ncs0.069580.0501
1019AX-RAY DIFFRACTIONLocal ncs0.060270.0501
1020AX-RAY DIFFRACTIONLocal ncs0.060270.0501
1121AX-RAY DIFFRACTIONLocal ncs0.052570.0501
1122AX-RAY DIFFRACTIONLocal ncs0.052570.0501
1223AX-RAY DIFFRACTIONLocal ncs0.05790.0501
1224AX-RAY DIFFRACTIONLocal ncs0.05790.0501
1325AX-RAY DIFFRACTIONLocal ncs0.054670.0501
1326AX-RAY DIFFRACTIONLocal ncs0.054670.0501
1427AX-RAY DIFFRACTIONLocal ncs0.056890.0501
1428AX-RAY DIFFRACTIONLocal ncs0.056890.0501
1529AX-RAY DIFFRACTIONLocal ncs0.058260.0501
1530AX-RAY DIFFRACTIONLocal ncs0.058260.0501
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
1.9-1.9490.3566360.349115640.349122330.8830.89399.73020.35
1.949-2.0030.3325880.32113990.321119950.9010.90799.93330.318
2.003-2.0610.3256320.289110360.291116760.9170.92999.93150.289
2.061-2.1240.2735690.258107590.259113330.9440.94799.95590.253
2.124-2.1940.2715720.223103950.226109670.9430.9621000.213
2.194-2.2710.2244660.2101600.201106300.9630.97199.96240.185
2.271-2.3560.2355150.18697580.189102810.9620.97699.92220.168
2.356-2.4520.2324710.16494070.16798820.9650.98399.95950.144
2.452-2.5610.2184670.15590590.15895280.9720.98699.9790.136
2.561-2.6860.2034570.15286420.15590990.9750.9871000.132
2.686-2.8310.2054130.15282400.15586540.9740.98699.98840.132
2.831-3.0030.2014080.15678050.15982150.9750.98599.97570.138
3.003-3.210.1953970.1673190.16277170.9750.98599.9870.146
3.21-3.4670.2033490.16368570.16572070.9760.98599.98610.154
3.467-3.7970.1993340.15563430.15766770.9790.9881000.149
3.797-4.2440.1483140.1257430.12160570.9870.9921000.119
4.244-4.8990.1382560.11251030.11453590.9890.9931000.116
4.899-5.9940.1972270.14443540.14745820.980.9999.97820.146
5.994-8.4560.1851920.14734030.14935990.9840.98999.88890.152
8.456-83.7410.1711030.17520080.17521120.9840.97699.95270.187
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)
11.5948-0.09560.25450.88670.20811.34360.0252-0.1776-0.03570.1075-0.0136-0.04940.0090.1456-0.01160.0143-0.0093-0.00740.0490.0110.012131.9955.4698-25.3981
21.4879-0.03490.16021.30210.11461.1429-0.00930.09990.2436-0.0988-0.03340.1461-0.1963-0.15210.04270.0410.0293-0.02410.03070.00830.1073-12.424727.3068-57.1476
31.8257-0.0302-0.04711.1643-0.03671.1075-0.015-0.3342-0.10430.1875-0.01810.12130.0709-0.18310.03310.04-0.01730.03140.09710.00480.0375-9.19361.0062-22.8086
41.36840.1416-0.10211.0192-0.01071.55930.03280.1398-0.2674-0.1012-0.01870.14450.2657-0.2427-0.01410.0951-0.0496-0.01480.0589-0.03950.1035-11.2671-15.865-62.5679
51.47620.14560.13381.04760.10471.21310.00250.23350.2886-0.1664-0.002-0.1334-0.18960.1908-0.00050.0577-0.02810.02670.07890.06560.122128.723227.8399-62.325
61.32660.12270.00751.1346-0.26131.27320.04360.2234-0.336-0.1496-0.0298-0.22060.30370.2735-0.01390.11890.06530.03030.0988-0.06950.151530.2504-15.5723-63.5525
Refinement TLS groupSelection: ALL

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