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- PDB-6o1j: Alpha-L-fucosidase AlfC fucosyltransferase mutant N243A -

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Basic information

Entry
Database: PDB / ID: 6o1j
TitleAlpha-L-fucosidase AlfC fucosyltransferase mutant N243A
ComponentsAlfC
KeywordsHYDROLASE / Fucosidase / fucosyltransferase / AlfC
Function / homology
Function and homology information


alpha-L-fucosidase activity / fucose metabolic process
Similarity search - Function
Alpha-L-fucosidase, metazoa-type / Glycoside hydrolase, family 29 / Alpha-L-fucosidase / Alpha-L-fucosidase / Glycoside hydrolase superfamily
Similarity search - Domain/homology
beta-L-fucopyranose / Alpha-L-fucosidase / Alpha-L-fucosidase
Similarity search - Component
Biological speciesLactobacillus casei (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / molecular replacement / Resolution: 2 Å
AuthorsKlontz, E.H. / Sundberg, E.J.
CitationJournal: Nat Commun / Year: 2020
Title: Structure and dynamics of an alpha-fucosidase reveal a mechanism for highly efficient IgG transfucosylation.
Authors: Klontz, E.H. / Li, C. / Kihn, K. / Fields, J.K. / Beckett, D. / Snyder, G.A. / Wintrode, P.L. / Deredge, D. / Wang, L.X. / Sundberg, E.J.
History
DepositionFeb 20, 2019Deposition site: RCSB / Processing site: RCSB
Revision 1.0Feb 26, 2020Provider: repository / Type: Initial release
Revision 1.1Jul 29, 2020Group: Data collection / Derived calculations / Structure summary
Category: chem_comp / entity ...chem_comp / entity / pdbx_chem_comp_identifier / pdbx_entity_nonpoly / struct_site / struct_site_gen
Item: _chem_comp.name / _entity.pdbx_description / _pdbx_entity_nonpoly.name
Description: Carbohydrate remediation / Provider: repository / Type: Remediation
Revision 1.2Dec 16, 2020Group: Database references / Structure summary / Category: chem_comp / citation / citation_author
Item: _chem_comp.pdbx_synonyms / _citation.country ..._chem_comp.pdbx_synonyms / _citation.country / _citation.journal_abbrev / _citation.journal_id_CSD / _citation.journal_id_ISSN / _citation.journal_volume / _citation.page_first / _citation.page_last / _citation.pdbx_database_id_DOI / _citation.pdbx_database_id_PubMed / _citation.title / _citation.year
Revision 1.3Oct 11, 2023Group: Data collection / Database references / Refinement description
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / pdbx_initial_refinement_model / struct_ncs_dom_lim
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession ..._database_2.pdbx_DOI / _database_2.pdbx_database_accession / _struct_ncs_dom_lim.beg_auth_comp_id / _struct_ncs_dom_lim.beg_label_asym_id / _struct_ncs_dom_lim.beg_label_comp_id / _struct_ncs_dom_lim.beg_label_seq_id / _struct_ncs_dom_lim.end_auth_comp_id / _struct_ncs_dom_lim.end_label_asym_id / _struct_ncs_dom_lim.end_label_comp_id / _struct_ncs_dom_lim.end_label_seq_id

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: AlfC
B: AlfC
C: AlfC
D: AlfC
E: AlfC
F: AlfC
G: AlfC
H: AlfC
hetero molecules


Theoretical massNumber of molelcules
Total (without water)315,92716
Polymers314,6148
Non-polymers1,3138
Water13,709761
1
A: AlfC
C: AlfC
E: AlfC
G: AlfC
hetero molecules


Theoretical massNumber of molelcules
Total (without water)157,9648
Polymers157,3074
Non-polymers6574
Water724
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
2
B: AlfC
H: AlfC
hetero molecules

D: AlfC
F: AlfC
hetero molecules


Theoretical massNumber of molelcules
Total (without water)157,9648
Polymers157,3074
Non-polymers6574
Water724
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
crystal symmetry operation1_455x-1,y,z1
3
D: AlfC
F: AlfC
hetero molecules

B: AlfC
H: AlfC
hetero molecules


Theoretical massNumber of molelcules
Total (without water)157,9648
Polymers157,3074
Non-polymers6574
Water724
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
crystal symmetry operation1_655x+1,y,z1
Unit cell
Length a, b, c (Å)80.301, 259.921, 80.350
Angle α, β, γ (deg.)90.000, 118.810, 90.000
Int Tables number4
Space group name H-MP1211
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11A
21B
12A
22C
13A
23D
14A
24E
15A
25F
16A
26G
17A
27H
18B
28C
19B
29D
110B
210E
111B
211F
112B
212G
113B
213H
114C
214D
115C
215E
116C
216F
117C
217G
118C
218H
119D
219E
120D
220F
121D
221G
122D
222H
123E
223F
124E
224G
125E
225H
126F
226G
127F
227H
128G
228H

NCS domain segments:

Component-ID: 0 / Refine code: 0

Dom-IDEns-IDBeg auth comp-IDBeg label comp-IDEnd auth comp-IDEnd label comp-IDAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
11METMETLEULEUAA1 - 3451 - 345
21METMETLEULEUBB1 - 3451 - 345
12ASNASNLEULEUAA2 - 3452 - 345
22ASNASNLEULEUCC2 - 3452 - 345
13METMETLEULEUAA1 - 3451 - 345
23METMETLEULEUDD1 - 3451 - 345
14ASNASNLEULEUAA2 - 3452 - 345
24ASNASNLEULEUEE2 - 3452 - 345
15METMETLEULEUAA1 - 3451 - 345
25METMETLEULEUFF1 - 3451 - 345
16ASNASNLEULEUAA4 - 3454 - 345
26ASNASNLEULEUGG4 - 3454 - 345
17METMETLEULEUAA1 - 3451 - 345
27METMETLEULEUHH1 - 3451 - 345
18ASNASNARGARGBB2 - 3442 - 344
28ASNASNARGARGCC2 - 3442 - 344
19METMETLEULEUBB1 - 3451 - 345
29METMETLEULEUDD1 - 3451 - 345
110ASNASNLEULEUBB2 - 3452 - 345
210ASNASNLEULEUEE2 - 3452 - 345
111METMETLEULEUBB1 - 3451 - 345
211METMETLEULEUFF1 - 3451 - 345
112ASNASNLEULEUBB4 - 3454 - 345
212ASNASNLEULEUGG4 - 3454 - 345
113METMETLEULEUBB1 - 3451 - 345
213METMETLEULEUHH1 - 3451 - 345
114ASNASNLEULEUCC2 - 3452 - 345
214ASNASNLEULEUDD2 - 3452 - 345
115ASNASNLEULEUCC2 - 3452 - 345
215ASNASNLEULEUEE2 - 3452 - 345
116ASNASNLEULEUCC2 - 3452 - 345
216ASNASNLEULEUFF2 - 3452 - 345
117ASNASNLEULEUCC4 - 3454 - 345
217ASNASNLEULEUGG4 - 3454 - 345
118ASNASNLEULEUCC2 - 3452 - 345
218ASNASNLEULEUHH2 - 3452 - 345
119ASNASNLEULEUDD2 - 3452 - 345
219ASNASNLEULEUEE2 - 3452 - 345
120METMETLEULEUDD1 - 3451 - 345
220METMETLEULEUFF1 - 3451 - 345
121ASNASNLEULEUDD4 - 3454 - 345
221ASNASNLEULEUGG4 - 3454 - 345
122METMETLEULEUDD1 - 3451 - 345
222METMETLEULEUHH1 - 3451 - 345
123ASNASNLEULEUEE2 - 3452 - 345
223ASNASNLEULEUFF2 - 3452 - 345
124ASNASNLEULEUEE4 - 3454 - 345
224ASNASNLEULEUGG4 - 3454 - 345
125ASNASNLEULEUEE2 - 3452 - 345
225ASNASNLEULEUHH2 - 3452 - 345
126ASNASNLEULEUFF4 - 3454 - 345
226ASNASNLEULEUGG4 - 3454 - 345
127METMETLEULEUFF1 - 3451 - 345
227METMETLEULEUHH1 - 3451 - 345
128ASNASNLEULEUGG4 - 3454 - 345
228ASNASNLEULEUHH4 - 3454 - 345

NCS ensembles :
ID
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28

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Components

#1: Protein
AlfC


Mass: 39326.727 Da / Num. of mol.: 8 / Mutation: N243A
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Lactobacillus casei (bacteria) / Production host: Escherichia coli BL21(DE3) (bacteria)
References: UniProt: K0NB39, UniProt: A0A422MHI3*PLUS, alpha-L-fucosidase
#2: Sugar
ChemComp-FUL / beta-L-fucopyranose / beta-L-fucose / 6-deoxy-beta-L-galactopyranose / L-fucose / fucose / 6-DEOXY-BETA-L-GALACTOSE / Fucose


Type: L-saccharide, beta linking / Mass: 164.156 Da / Num. of mol.: 8
Source method: isolated from a genetically manipulated source
Formula: C6H12O5
IdentifierTypeProgram
LFucpbCONDENSED IUPAC CARBOHYDRATE SYMBOLGMML 1.0
b-L-fucopyranoseCOMMON NAMEGMML 1.0
b-L-FucpIUPAC CARBOHYDRATE SYMBOLPDB-CARE 1.0
FucSNFG CARBOHYDRATE SYMBOLGMML 1.0
#3: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 761 / Source method: isolated from a natural source / Formula: H2O

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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: 2.34 Å3/Da / Density % sol: 47.33 %
Crystal growTemperature: 298 K / Method: vapor diffusion, sitting drop / pH: 7
Details: 250 nL 20 mg/mL AlfC + 250 nL mother liquor (18% w/v PEG3350, 0.1 M Bis-Tris propane, pH 7, 20 mM sodium/potassium phosphate, 1% v/v glycerol), crystals appeared after five days, harvested ...Details: 250 nL 20 mg/mL AlfC + 250 nL mother liquor (18% w/v PEG3350, 0.1 M Bis-Tris propane, pH 7, 20 mM sodium/potassium phosphate, 1% v/v glycerol), crystals appeared after five days, harvested in mother liquor + 20% v/v glycerol + 5 mM 4-NP-Fuc for ~1 minute, flash-cooled in liquid nitrogen

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Data collection

DiffractionMean temperature: 100 K / Serial crystal experiment: N
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 23-ID-B / Wavelength: 1.03321 Å
DetectorType: DECTRIS EIGER X 16M / Detector: PIXEL / Date: Mar 5, 2018
RadiationMonochromator: Double crystal cryo-cooled Si(111) / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 1.03321 Å / Relative weight: 1
Reflection twin
Crystal-IDIDOperatorDomain-IDFraction
11H, K, L10.645
11-L, -K, -H20.355
ReflectionResolution: 2→47.75 Å / Num. obs: 177005 / % possible obs: 91.5 % / Redundancy: 2.3 % / CC1/2: 0.998 / Rmerge(I) obs: 0.055 / Rpim(I) all: 0.042 / Rrim(I) all: 0.07 / Net I/σ(I): 8.5
Reflection shell

Diffraction-ID: 1

Resolution (Å)Redundancy (%)Rmerge(I) obsNum. measured allNum. unique obsCC1/2Rpim(I) allRrim(I) allNet I/σ(I) obs% possible all
2-2.031.80.827987255590.4620.6751.0740.758.4
10.95-47.752.50.016292811730.9990.0120.0212696.4

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Phasing

PhasingMethod: molecular replacement

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Processing

Software
NameVersionClassificationNB
XDSdata reduction
Aimless0.7.3data scaling
MOLREPphasing
REFMAC5.8.0238refinement
PDB_EXTRACT3.24data extraction
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: PDB entry 6O18
Resolution: 2→47.75 Å / Cor.coef. Fo:Fc: 0.963 / Cor.coef. Fo:Fc free: 0.957 / SU B: 7.235 / SU ML: 0.106 / Cross valid method: THROUGHOUT / σ(F): 0 / ESU R: 0.053 / ESU R Free: 0.039
Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS U VALUES : WITH TLS ADDED
RfactorNum. reflection% reflectionSelection details
Rfree0.2426 9045 5.1 %RANDOM
Rwork0.23 ---
obs0.2307 167908 91.28 %-
Solvent computationIon probe radii: 0.7 Å / Shrinkage radii: 0.7 Å / VDW probe radii: 1 Å
Displacement parametersBiso max: 122.2 Å2 / Biso mean: 48.55 Å2 / Biso min: 24.53 Å2
Baniso -1Baniso -2Baniso -3
1--4.9 Å2-0 Å219.73 Å2
2---9.95 Å20 Å2
3---14.84 Å2
Refinement stepCycle: final / Resolution: 2→47.75 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms20791 0 88 761 21640
Biso mean--55.73 48.19 -
Num. residues----2590
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0080.01321454
X-RAY DIFFRACTIONr_bond_other_d0.0030.01718580
X-RAY DIFFRACTIONr_angle_refined_deg1.461.64429146
X-RAY DIFFRACTIONr_angle_other_deg1.3371.58343274
X-RAY DIFFRACTIONr_dihedral_angle_1_deg6.94252572
X-RAY DIFFRACTIONr_dihedral_angle_2_deg34.86824.2641175
X-RAY DIFFRACTIONr_dihedral_angle_3_deg15.405153383
X-RAY DIFFRACTIONr_dihedral_angle_4_deg8.5521564
X-RAY DIFFRACTIONr_chiral_restr0.0740.22679
X-RAY DIFFRACTIONr_gen_planes_refined0.0070.0224314
X-RAY DIFFRACTIONr_gen_planes_other0.0020.024595
Refine LS restraints NCS

Refine-ID: X-RAY DIFFRACTION / Type: interatomic distance / Weight position: 0.05

Ens-IDDom-IDAuth asym-IDNumberRms dev position (Å)
11A111600.08
12B111600.08
21A111470.08
22C111470.08
31A111650.08
32D111650.08
41A113010.08
42E113010.08
51A111590.08
52F111590.08
61A109700.08
62G109700.08
71A111460.08
72H111460.08
81B111950.06
82C111950.06
91B112090.07
92D112090.07
101B112190.06
102E112190.06
111B111300.07
112F111300.07
121B109310.07
122G109310.07
131B111170.07
132H111170.07
141C112580.06
142D112580.06
151C112000.07
152E112000.07
161C112120.07
162F112120.07
171C110550.07
172G110550.07
181C111390.07
182H111390.07
191D112330.07
192E112330.07
201D112050.07
202F112050.07
211D109870.07
212G109870.07
221D112110.07
222H112110.07
231E111210.08
232F111210.08
241E110150.07
242G110150.07
251E112300.07
252H112300.07
261F110480.07
262G110480.07
271F111720.07
272H111720.07
281G109610.07
282H109610.07
LS refinement shellResolution: 1.999→2.051 Å / Rfactor Rfree error: 0 / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree1.381 389 -
Rwork1.187 7904 -
all-8293 -
obs--57.93 %
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)
11.36210.4246-0.10371.51110.37520.90610.0926-0.09950.17640.2737-0.1260.2682-0.0445-0.12570.03350.0752-0.01440.01790.2459-0.01060.1743-21.4820.2643.27
21.18330.0805-0.19911.2945-0.14290.67290.0111-0.09150.03060.1533-0.0246-0.21060.00660.13590.01340.02990.0047-0.07620.2663-0.00290.2784-4.53640.61917.819
30.92780.6383-0.10941.9306-0.24011.3307-0.15820.262-0.2686-0.43240.1004-0.10650.314-0.01420.05780.1591-0.04-0.00510.3588-0.07320.2576-17.458-29.404-23.375
41.2391-0.196-0.3541.22020.28290.88050.03590.30390.0077-0.1794-0.02730.2169-0.0447-0.1438-0.00870.0369-0.0023-0.08410.33290.01470.312839.74244.7320.757
50.91480.2868-0.18452.0952-1.09651.8601-0.13290.1344-0.2252-0.3963-0.0978-0.68220.34250.21170.23060.10260.02710.0970.31570.00070.390118.806-9.675-23.243
61.0982-0.2682-0.19251.5-0.11950.65290.07780.28570.0886-0.2662-0.0885-0.0124-0.0481-0.06640.01080.07040.0044-0.04350.31790.03490.267657.70982.267-1.674
71.27990.28510.16861.3881-0.56151.3345-0.04730.15310.3068-0.06340.08830.1901-0.2863-0.188-0.04090.14220.0354-0.03850.36550.060.2536-7.95618.678-30.809
81.16640.49-0.10611.1882-0.01140.80920.1289-0.4004-0.02750.4775-0.1539-0.0683-0.05950.10690.02490.2376-0.0691-0.04910.39390.01170.3165-21.66372.55737.103
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1A1 - 345
2X-RAY DIFFRACTION2B1 - 345
3X-RAY DIFFRACTION3C2 - 345
4X-RAY DIFFRACTION4D1 - 345
5X-RAY DIFFRACTION5E2 - 345
6X-RAY DIFFRACTION6F1 - 345
7X-RAY DIFFRACTION7G1 - 345
8X-RAY DIFFRACTION8H1 - 345

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