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- PDB-6o1c: Alpha-L-fucosidase AlfC D200A mutant in complex with 4-nitropheny... -

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Basic information

Entry
Database: PDB / ID: 6o1c
TitleAlpha-L-fucosidase AlfC D200A mutant in complex with 4-nitrophenyl-a-L-fucopyranoside substrate
ComponentsAlfC
KeywordsHYDROLASE / Fucosidase / substrate / 4NP-Fuc / 4-nitrophenyl-a-L-fucopyranoside
Function / homology
Function and homology information


alpha-L-fucosidase activity / fucose metabolic process / glycoside catabolic process / lysosome
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
4-nitrophenyl 6-deoxy-alpha-L-galactopyranoside / Uncharacterized protein / Uncharacterized protein
Similarity search - Component
Biological speciesLactobacillus casei (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / molecular replacement / Resolution: 2.6 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 19, 2019Deposition site: RCSB / Processing site: RCSB
Revision 1.0Feb 19, 2020Provider: repository / Type: Initial release
Revision 2.0Jul 29, 2020Group: Atomic model / Derived calculations / Structure summary
Category: atom_site / chem_comp ...atom_site / chem_comp / entity / pdbx_entity_nonpoly / struct_site / struct_site_gen
Item: _atom_site.auth_atom_id / _atom_site.label_atom_id ..._atom_site.auth_atom_id / _atom_site.label_atom_id / _chem_comp.mon_nstd_flag / _chem_comp.name / _chem_comp.type / _entity.pdbx_description / _pdbx_entity_nonpoly.name
Description: Carbohydrate remediation / Provider: repository / Type: Remediation
Revision 2.1Dec 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 2.2Oct 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)316,88816
Polymers314,6068
Non-polymers2,2828
Water4,179232
1
A: AlfC
D: AlfC
E: AlfC
G: AlfC
hetero molecules


Theoretical massNumber of molelcules
Total (without water)158,4448
Polymers157,3034
Non-polymers1,1414
Water724
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
2
B: AlfC
C: AlfC
F: AlfC
H: AlfC
hetero molecules


Theoretical massNumber of molelcules
Total (without water)158,4448
Polymers157,3034
Non-polymers1,1414
Water724
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Unit cell
Length a, b, c (Å)89.734, 139.079, 263.373
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
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: _ / Refine code: _

Dom-IDEns-IDBeg auth comp-IDBeg label comp-IDEnd auth comp-IDEnd label comp-IDAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
11ASNASNARGARGAA2 - 3442 - 344
21ASNASNARGARGBB2 - 3442 - 344
12ASNASNLEULEUAA2 - 3452 - 345
22ASNASNLEULEUCC2 - 3452 - 345
13ASNASNLEULEUAA2 - 3452 - 345
23ASNASNLEULEUDD2 - 3452 - 345
14ASPASPLEULEUAA3 - 3453 - 345
24ASPASPLEULEUEE3 - 3453 - 345
15ASPASPARGARGAA3 - 3443 - 344
25ASPASPARGARGFF3 - 3443 - 344
16ASPASPLEULEUAA3 - 3453 - 345
26ASPASPLEULEUGG3 - 3453 - 345
17VALVALLEULEUAA5 - 3455 - 345
27VALVALLEULEUHH5 - 3455 - 345
18ASNASNASNASNBB2 - 3432 - 343
28ASNASNASNASNCC2 - 3432 - 343
19ASNASNARGARGBB2 - 3442 - 344
29ASNASNARGARGDD2 - 3442 - 344
110ASPASPASNASNBB3 - 3433 - 343
210ASPASPASNASNEE3 - 3433 - 343
111ASPASPARGARGBB3 - 3443 - 344
211ASPASPARGARGFF3 - 3443 - 344
112ASPASPASNASNBB3 - 3433 - 343
212ASPASPASNASNGG3 - 3433 - 343
113VALVALARGARGBB5 - 3445 - 344
213VALVALARGARGHH5 - 3445 - 344
114ASNASNLEULEUCC2 - 3452 - 345
214ASNASNLEULEUDD2 - 3452 - 345
115ASPASPLEULEUCC3 - 3453 - 345
215ASPASPLEULEUEE3 - 3453 - 345
116ASPASPASNASNCC3 - 3433 - 343
216ASPASPASNASNFF3 - 3433 - 343
117ASPASPLEULEUCC3 - 3453 - 345
217ASPASPLEULEUGG3 - 3453 - 345
118VALVALLEULEUCC5 - 3455 - 345
218VALVALLEULEUHH5 - 3455 - 345
119ASPASPLEULEUDD3 - 3453 - 345
219ASPASPLEULEUEE3 - 3453 - 345
120ASPASPARGARGDD3 - 3443 - 344
220ASPASPARGARGFF3 - 3443 - 344
121ASPASPLEULEUDD3 - 3453 - 345
221ASPASPLEULEUGG3 - 3453 - 345
122VALVALLEULEUDD5 - 3455 - 345
222VALVALLEULEUHH5 - 3455 - 345
123ASPASPASNASNEE3 - 3433 - 343
223ASPASPASNASNFF3 - 3433 - 343
124ASPASPLEULEUEE3 - 3453 - 345
224ASPASPLEULEUGG3 - 3453 - 345
125VALVALLEULEUEE5 - 3455 - 345
225VALVALLEULEUHH5 - 3455 - 345
126ASPASPARGARGFF3 - 3443 - 344
226ASPASPARGARGGG3 - 3443 - 344
127VALVALARGARGFF5 - 3445 - 344
227VALVALARGARGHH5 - 3445 - 344
128VALVALLEULEUGG5 - 3455 - 345
228VALVALLEULEUHH5 - 3455 - 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: 39325.742 Da / Num. of mol.: 8 / Mutation: D200A
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-JFZ / 4-nitrophenyl 6-deoxy-alpha-L-galactopyranoside / 4-nitrophenyl-alpha-L-fucose / 4-nitrophenyl 6-deoxy-alpha-L-galactoside / 4-nitrophenyl 6-deoxy-L-galactoside / 4-nitrophenyl 6-deoxy-galactoside


Type: L-saccharide / Mass: 285.250 Da / Num. of mol.: 8 / Source method: obtained synthetically / Formula: C12H15NO7 / Feature type: SUBJECT OF INVESTIGATION
#3: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 232 / 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: 2.61 Å3/Da / Density % sol: 52.91 %
Crystal growTemperature: 298 K / Method: vapor diffusion, sitting drop / pH: 7
Details: 250 nL 20 mg/mL AlfC D200Aof + 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, ...Details: 250 nL 20 mg/mL AlfC D200Aof + 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, were harvested in mother liquor + 20% v/v glycerol + 5 mM 4NP-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: SSRL / Beamline: BL12-2 / Wavelength: 1.03317 Å
DetectorType: DECTRIS PILATUS3 6M / Detector: PIXEL / Date: Jan 14, 2019
RadiationMonochromator: double crystal Si(111) / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 1.03317 Å / Relative weight: 1
ReflectionResolution: 2.6→39.43 Å / Num. obs: 97990 / % possible obs: 96.7 % / Redundancy: 3.4 % / CC1/2: 0.994 / Rmerge(I) obs: 0.109 / Rpim(I) all: 0.068 / Rrim(I) all: 0.129 / Net I/σ(I): 6.3 / Num. measured all: 330144 / Scaling rejects: 6
Reflection shell

Diffraction-ID: 1 / Redundancy: 3.3 %

Resolution (Å)Rmerge(I) obsNum. unique obsCC1/2Rpim(I) allRrim(I) all% possible all
2.6-2.641.36749330.5820.8551.62198.5
14.24-39.430.0546150.9970.0350.06586.6

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Phasing

PhasingMethod: molecular replacement

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Processing

Software
NameVersionClassificationNB
XDSdata reduction
Aimless0.7.1data scaling
MOLREPphasing
REFMAC5.8.0238refinement
PDB_EXTRACT3.24data extraction
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: PDB entry 6O18
Resolution: 2.6→39.43 Å / Cor.coef. Fo:Fc: 0.944 / Cor.coef. Fo:Fc free: 0.934 / SU B: 36.953 / SU ML: 0.34 / Cross valid method: THROUGHOUT / σ(F): 0 / ESU R: 0.954 / ESU R Free: 0.333
Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS U VALUES : WITH TLS ADDED
RfactorNum. reflection% reflectionSelection details
Rfree0.2607 4998 5.1 %RANDOM
Rwork0.2315 ---
obs0.233 92904 95.82 %-
Solvent computationIon probe radii: 0.7 Å / Shrinkage radii: 0.7 Å / VDW probe radii: 1 Å
Displacement parametersBiso max: 146.92 Å2 / Biso mean: 70.728 Å2 / Biso min: 38.54 Å2
Baniso -1Baniso -2Baniso -3
1--2.38 Å2-0 Å20 Å2
2--3.03 Å2-0 Å2
3----0.65 Å2
Refinement stepCycle: final / Resolution: 2.6→39.43 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms20562 0 160 232 20954
Biso mean--77.81 62.29 -
Num. residues----2584
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0070.01321363
X-RAY DIFFRACTIONr_bond_other_d0.0030.01718283
X-RAY DIFFRACTIONr_angle_refined_deg1.3851.64629072
X-RAY DIFFRACTIONr_angle_other_deg1.3111.58142535
X-RAY DIFFRACTIONr_dihedral_angle_1_deg6.61352574
X-RAY DIFFRACTIONr_dihedral_angle_2_deg35.25424.1681154
X-RAY DIFFRACTIONr_dihedral_angle_3_deg15.609153244
X-RAY DIFFRACTIONr_dihedral_angle_4_deg7.8281563
X-RAY DIFFRACTIONr_chiral_restr0.0660.22674
X-RAY DIFFRACTIONr_gen_planes_refined0.0060.0224312
X-RAY DIFFRACTIONr_gen_planes_other0.0010.024630
Refine LS restraints NCS

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

Ens-IDDom-IDAuth asym-IDNumberRms dev position (Å)
11A109470.07
12B109470.07
21A108900.08
22C108900.08
31A111790.05
32D111790.05
41A108540.07
42E108540.07
51A107100.08
52F107100.08
61A107050.07
62G107050.07
71A104580.09
72H104580.09
81B108980.06
82C108980.06
91B108420.07
92D108420.07
101B106790.07
102E106790.07
111B107950.06
112F107950.06
121B105890.06
122G105890.06
131B104960.08
132H104960.08
141C109010.06
142D109010.06
151C107140.08
152E107140.08
161C107500.07
162F107500.07
171C105980.07
172G105980.07
181C106130.07
182H106130.07
191D108110.06
192E108110.06
201D107000.08
202F107000.08
211D106420.07
212G106420.07
221D104990.08
222H104990.08
231E107120.08
232F107120.08
241E106270.07
242G106270.07
251E104980.09
252H104980.09
261F104980.07
262G104980.07
271F105350.08
272H105350.08
281G103460.08
282H103460.08
LS refinement shellResolution: 2.6→2.667 Å / Rfactor Rfree error: 0 / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.42 368 -
Rwork0.393 6954 -
all-7322 -
obs--97.85 %
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)
12.1945-0.2286-0.43421.5164-0.21681.41-0.03570.22650.2947-0.09460.12680.1155-0.13490.0397-0.09110.8572-0.02740.00850.04810.04450.05456.249-44.659-5.887
21.2756-0.3360.21951.6448-0.01352.44460.0942-0.1889-0.06510.1615-0.0445-0.20970.20680.4647-0.04980.8462-0.0435-0.02640.14160.01620.036932.514-52.781-43.392
31.24390.46080.3271.671-0.47022.1680.11060.1261-0.0635-0.29320.01850.19660.4284-0.3201-0.1291.0061-0.02-0.05220.0854-0.00540.035911.98-55.324-89.106
42.30570.61640.63521.63620.05481.68270.1519-0.1353-0.38550.27190.0684-0.05990.43080.0489-0.22030.99530.085-0.06290.02490.01640.07898.357-93.6564.345
51.58890.03460.3721.37760.28731.7362-0.06770.37580.2039-0.23060.07530.4556-0.1309-0.0615-0.00770.9419-0.0696-0.11550.11160.08680.2002-12.55-76.223-25.675
61.3360.1256-0.03911.48070.18242.08480.04010.11470.14960.0503-0.054-0.4634-0.16230.83140.0140.9359-0.07680.03420.35770.06560.177843.538-32.634-78.407
72.1080.1025-0.14121.32620.21191.1095-0.1697-0.5372-0.32080.43280.11350.4390.304-0.06960.05621.09460.14250.25220.18020.12730.254-13.942-64.28923.161
81.3276-0.22380.27241.6907-0.55052.15150.07-0.0920.0378-0.05780.22950.5833-0.1943-0.9893-0.29940.95520.07050.03520.48510.12540.2227-1.542-34.617-55.789
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1A2 - 345
2X-RAY DIFFRACTION2B2 - 345
3X-RAY DIFFRACTION3C2 - 345
4X-RAY DIFFRACTION4D2 - 345
5X-RAY DIFFRACTION5E3 - 345
6X-RAY DIFFRACTION6F3 - 344
7X-RAY DIFFRACTION7G3 - 345
8X-RAY DIFFRACTION8H3 - 345

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