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- PDB-5f7s: Cycloalternan-degrading enzyme from Trueperella pyogenes -

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Basic information

Entry
Database: PDB / ID: 5f7s
TitleCycloalternan-degrading enzyme from Trueperella pyogenes
ComponentsGlycoside hydrolase family 31
KeywordsHYDROLASE / Structural Genomics / Center for Structural Genomics of Infectious Diseases / CSGID
Function / homology
Function and homology information


hydrolase activity, hydrolyzing O-glycosyl compounds / carbohydrate binding / carbohydrate metabolic process
Similarity search - Function
Glycoside hydrolase family 31, N-terminal domain / Glycosyl hydrolase 31 N-terminal galactose mutarotase-like domain / Glycoside hydrolase family 31 / Glycosyl hydrolases family 31 TIM-barrel domain / Galactose mutarotase-like domain superfamily / Golgi alpha-mannosidase II / Glycosyl hydrolase, all-beta / Glycosidases / Glycoside hydrolase superfamily / TIM Barrel ...Glycoside hydrolase family 31, N-terminal domain / Glycosyl hydrolase 31 N-terminal galactose mutarotase-like domain / Glycoside hydrolase family 31 / Glycosyl hydrolases family 31 TIM-barrel domain / Galactose mutarotase-like domain superfamily / Golgi alpha-mannosidase II / Glycosyl hydrolase, all-beta / Glycosidases / Glycoside hydrolase superfamily / TIM Barrel / Alpha-Beta Barrel / Immunoglobulin-like fold / Immunoglobulin-like / Sandwich / Mainly Beta / Alpha Beta
Similarity search - Domain/homology
DI(HYDROXYETHYL)ETHER / TRIETHYLENE GLYCOL / Glycoside hydrolase family 31
Similarity search - Component
Biological speciesTrueperella pyogenes (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.3 Å
AuthorsLight, S.H. / Anderson, W.F. / Center for Structural Genomics of Infectious Diseases (CSGID)
CitationJournal: Nat Microbiol / Year: 2016
Title: Structure to function of an alpha-glucan metabolic pathway that promotes Listeria monocytogenes pathogenesis.
Authors: Light, S.H. / Cahoon, L.A. / Halavaty, A.S. / Freitag, N.E. / Anderson, W.F.
History
DepositionDec 8, 2015Deposition site: RCSB / Processing site: RCSB
Revision 1.0Dec 30, 2015Provider: repository / Type: Initial release
Revision 1.1Nov 23, 2016Group: Database references
Revision 1.2Sep 27, 2023Group: Data collection / Database references ...Data collection / Database references / Derived calculations / Refinement description
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / pdbx_initial_refinement_model / pdbx_struct_conn_angle / struct_conn
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession ..._database_2.pdbx_DOI / _database_2.pdbx_database_accession / _pdbx_struct_conn_angle.ptnr1_auth_asym_id / _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.ptnr1_symmetry / _pdbx_struct_conn_angle.ptnr2_auth_asym_id / _pdbx_struct_conn_angle.ptnr2_auth_seq_id / _pdbx_struct_conn_angle.ptnr2_label_asym_id / _pdbx_struct_conn_angle.ptnr3_auth_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.ptnr3_symmetry / _pdbx_struct_conn_angle.value / _struct_conn.pdbx_dist_value / _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.ptnr1_symmetry / _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 / _struct_conn.ptnr2_symmetry

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Glycoside hydrolase family 31
B: Glycoside hydrolase family 31
hetero molecules


Theoretical massNumber of molelcules
Total (without water)165,42414
Polymers164,6572
Non-polymers76712
Water11,836657
1
A: Glycoside hydrolase family 31
hetero molecules


Theoretical massNumber of molelcules
Total (without water)82,6758
Polymers82,3281
Non-polymers3477
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA
2
B: Glycoside hydrolase family 31
hetero molecules


Theoretical massNumber of molelcules
Total (without water)82,7496
Polymers82,3281
Non-polymers4215
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA
Unit cell
Length a, b, c (Å)197.126, 104.663, 89.030
Angle α, β, γ (deg.)90.00, 90.74, 90.00
Int Tables number5
Space group name H-MC121

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Components

#1: Protein Glycoside hydrolase family 31 /


Mass: 82328.406 Da / Num. of mol.: 2
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Trueperella pyogenes (bacteria) / Gene: CQ11_05330 / Production host: Escherichia coli BL21 (bacteria) / References: UniProt: X4QP62
#2: Chemical
ChemComp-CA / CALCIUM ION


Mass: 40.078 Da / Num. of mol.: 9 / Source method: obtained synthetically / Formula: Ca
#3: Chemical ChemComp-PEG / DI(HYDROXYETHYL)ETHER / Diethylene glycol


Mass: 106.120 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C4H10O3
#4: Chemical ChemComp-PGE / TRIETHYLENE GLYCOL / Polyethylene glycol


Mass: 150.173 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: C6H14O4
#5: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 657 / 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: 2.79 Å3/Da / Density % sol: 55.89 %
Crystal growTemperature: 300 K / Method: vapor diffusion, sitting drop
Details: Protein solution: 12.0 mg/ml, 0.5 M NaCl, 0.01 M Tris-HCl pH 8.3 Crystallization condition: PACT (Qiagen) C11: 0.2 M Calcium chloride, 0.1 m Hepes pH 7, and 25% (w/v) PEG 600

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 21-ID-G / Wavelength: 0.97856 Å
DetectorType: MARMOSAIC 300 mm CCD / Detector: CCD / Date: Oct 12, 2015
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.97856 Å / Relative weight: 1
ReflectionResolution: 2.3→30 Å / Num. obs: 80180 / % possible obs: 100 % / Redundancy: 3.8 % / Rmerge(I) obs: 0.074 / Net I/σ(I): 15.2
Reflection shellResolution: 2.3→2.34 Å / Redundancy: 3.8 % / Rmerge(I) obs: 0.564 / Mean I/σ(I) obs: 2.2 / % possible all: 100

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Processing

Software
NameVersionClassification
REFMAC5.8.0135refinement
HKL-3000data reduction
HKL-3000data scaling
PHASERphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 4KMQ
Resolution: 2.3→30 Å / Cor.coef. Fo:Fc: 0.962 / Cor.coef. Fo:Fc free: 0.94 / SU B: 12.291 / SU ML: 0.15 / Cross valid method: THROUGHOUT / ESU R: 0.258 / ESU R Free: 0.196 / Stereochemistry target values: MAXIMUM LIKELIHOOD / Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS
RfactorNum. reflection% reflectionSelection details
Rfree0.21122 4135 5.2 %RANDOM
Rwork0.16965 ---
obs0.1718 76044 99.72 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.2 Å / Solvent model: MASK
Displacement parametersBiso mean: 36.376 Å2
Baniso -1Baniso -2Baniso -3
1-0.41 Å2-0 Å20.45 Å2
2---0.28 Å20 Å2
3----0.14 Å2
Refinement stepCycle: LAST / Resolution: 2.3→30 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms11226 0 36 657 11919
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0110.01911611
X-RAY DIFFRACTIONr_bond_other_d0.0020.0210414
X-RAY DIFFRACTIONr_angle_refined_deg1.5261.92715836
X-RAY DIFFRACTIONr_angle_other_deg0.882323909
X-RAY DIFFRACTIONr_dihedral_angle_1_deg3.42751398
X-RAY DIFFRACTIONr_dihedral_angle_2_deg27.93222.987596
X-RAY DIFFRACTIONr_dihedral_angle_3_deg10.872151699
X-RAY DIFFRACTIONr_dihedral_angle_4_deg12.17715102
X-RAY DIFFRACTIONr_chiral_restr0.0980.21613
X-RAY DIFFRACTIONr_gen_planes_refined0.0070.02113342
X-RAY DIFFRACTIONr_gen_planes_other0.0010.022904
X-RAY DIFFRACTIONr_nbd_refined
X-RAY DIFFRACTIONr_nbd_other
X-RAY DIFFRACTIONr_nbtor_refined
X-RAY DIFFRACTIONr_nbtor_other
X-RAY DIFFRACTIONr_xyhbond_nbd_refined
X-RAY DIFFRACTIONr_xyhbond_nbd_other
X-RAY DIFFRACTIONr_metal_ion_refined
X-RAY DIFFRACTIONr_metal_ion_other
X-RAY DIFFRACTIONr_symmetry_vdw_refined
X-RAY DIFFRACTIONr_symmetry_vdw_other
X-RAY DIFFRACTIONr_symmetry_hbond_refined
X-RAY DIFFRACTIONr_symmetry_hbond_other
X-RAY DIFFRACTIONr_symmetry_metal_ion_refined
X-RAY DIFFRACTIONr_symmetry_metal_ion_other
X-RAY DIFFRACTIONr_mcbond_it1.1872.3645607
X-RAY DIFFRACTIONr_mcbond_other1.1852.3635606
X-RAY DIFFRACTIONr_mcangle_it1.9843.5397000
X-RAY DIFFRACTIONr_mcangle_other1.9853.547001
X-RAY DIFFRACTIONr_scbond_it1.4892.5816004
X-RAY DIFFRACTIONr_scbond_other1.4892.5816004
X-RAY DIFFRACTIONr_scangle_it
X-RAY DIFFRACTIONr_scangle_other2.5183.7978837
X-RAY DIFFRACTIONr_long_range_B_refined5.11119.79613446
X-RAY DIFFRACTIONr_long_range_B_other5.11119.79813447
X-RAY DIFFRACTIONr_rigid_bond_restr
X-RAY DIFFRACTIONr_sphericity_free
X-RAY DIFFRACTIONr_sphericity_bonded
LS refinement shellResolution: 2.3→2.359 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.271 293 -
Rwork0.243 5484 -
obs--97.6 %
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)
10.3341-1.34721.003212.0675-4.38713.0381-0.0042-0.03920.00570.10830.01010.263-0.036-0.136-0.00580.07130.00050.02510.1366-0.01520.1311465.412627.246540.0012
22.87772.1214-3.16942.4395-2.04374.79960.1429-0.1787-0.28350.31-0.3396-0.3503-0.35690.30960.19670.1308-0.0588-0.01570.15930.06790.1389473.638920.07943.2505
31.39371.6527-2.42522.5754-2.71484.681-0.1655-0.1504-0.3495-0.1329-0.2711-0.4050.36050.32150.43660.03750.02230.00820.17230.07190.2383476.125319.706738.5828
41.3664-0.3963-0.36071.87450.33820.5097-0.03630.0295-0.0544-0.0367-0.0078-0.20780.03880.09240.0440.03440.01230.02570.11480.01730.0454476.519746.280126.6487
52.4832-0.9437-0.33981.07070.43470.3655-0.04230.3409-0.156-0.0575-0.0422-0.04210.0348-0.03590.08450.1684-0.00360.04580.1897-0.00350.0985464.237351.658117.0044
61.80530.8550.01861.8923-0.11840.2308-0.04450.0423-0.14360.07670.05660.2971-0.0285-0.0945-0.01210.07110.02180.03390.1203-0.02360.1104433.687952.904632.3551
71.1258-0.2074-0.0723.435-0.36260.5419-0.06880.0938-0.29670.194-0.02190.39960.0179-0.12650.09060.0302-0.02060.03140.0759-0.03950.1501441.124840.581630.0617
85.968-4.581-1.15358.79724.07836.7766-0.1871-0.492-0.32920.27780.08540.25980.14750.14380.10170.02790.0050.00660.07010.04080.117453.027230.67334.0957
90.8784-0.0333-0.04961.15690.06830.517-0.00070.0377-0.0431-0.0094-0.0512-0.0422-0.0597-0.00630.05190.0560.0125-0.00090.10850.00050.0095455.522957.680429.2053
104.73790.1768-1.10383.8266-0.73431.29320.25020.21570.4274-0.0698-0.0958-0.3834-0.36160.1474-0.15440.1638-0.05950.02620.06120.01830.161467.103980.682828.7461
111.0642-0.1109-0.07672.8671-1.05281.3180.0518-0.0631-0.04970.1499-0.16090.0941-0.18940.08090.10910.1249-0.00620.04810.18650.04050.1108470.407226.0257-6.5573
121.66440.4254-3.51170.7932-0.02688.5261-0.1457-0.2318-0.19280.0806-0.0695-0.18050.43450.46910.21520.07890.02110.02420.09030.07410.1378475.310511.8068-1.0592
131.86941.6884-2.69842.7255-3.01175.4507-0.1186-0.1768-0.3253-0.1285-0.2639-0.26280.29970.33680.38250.05580.02360.02680.15460.09280.1485475.346217.8084-6.105
142.71521.64660.07865.79872.06071.4173-0.01650.1066-0.1243-0.3228-0.0513-0.4191-0.0050.12720.06770.09470.03130.05860.23470.02440.1677483.101139.1389-26.7685
151.1581-0.3965-0.25190.76380.1080.2775-0.03820.0463-0.05180.0384-0.038-0.05720.0066-0.00810.07620.0834-0.00210.01350.1184-0.00910.0353460.67250.581-18.4335
161.0114-0.4919-0.28841.14740.28410.7799-0.08240.0112-0.20510.196-0.05130.31510.035-0.22310.13370.0649-0.03740.04950.1449-0.0560.1229436.23844.8699-14.3158
177.1496-3.7411-3.36938.15826.72877.6595-0.1581-0.2664-0.13860.092-0.01970.21230.25130.04370.17790.01880.01420.01040.03480.03090.0493453.464930.4226-11.685
180.63160.07240.00710.78430.14230.5187-0.0004-0.0032-0.08220.0863-0.07050.0389-0.0058-0.07430.07080.070.00770.00830.1208-0.00770.0223452.154152.9532-14.1741
196.792-5.57832.73314.5834-2.26281.4091-0.2711-0.264-0.28430.24380.22260.2295-0.2748-0.17840.04860.15060.0145-0.02480.1024-0.04370.1333449.977175.4051-16.6572
201.8228-0.4482-0.59031.49540.16791.02770.09430.00230.296-0.0854-0.0613-0.3133-0.14370.0713-0.03290.077-0.01210.00630.07650.02570.0983466.617673.4398-19.8024
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1A1 - 17
2X-RAY DIFFRACTION2A18 - 50
3X-RAY DIFFRACTION3A51 - 144
4X-RAY DIFFRACTION4A145 - 254
5X-RAY DIFFRACTION5A255 - 300
6X-RAY DIFFRACTION6A301 - 378
7X-RAY DIFFRACTION7A379 - 416
8X-RAY DIFFRACTION8A417 - 432
9X-RAY DIFFRACTION9A433 - 694
10X-RAY DIFFRACTION10A695 - 730
11X-RAY DIFFRACTION11B1 - 38
12X-RAY DIFFRACTION12B39 - 53
13X-RAY DIFFRACTION13B54 - 138
14X-RAY DIFFRACTION14B139 - 172
15X-RAY DIFFRACTION15B173 - 352
16X-RAY DIFFRACTION16B353 - 416
17X-RAY DIFFRACTION17B417 - 431
18X-RAY DIFFRACTION18B432 - 626
19X-RAY DIFFRACTION19B627 - 631
20X-RAY DIFFRACTION20B632 - 730

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