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- PDB-7hk7: Crystal Structure of N-methylhydantoinase in complex with Mg-ADPNP -

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Basic information

Entry
Database: PDB / ID: 7hk7
TitleCrystal Structure of N-methylhydantoinase in complex with Mg-ADPNP
ComponentsN-methylhydantoinase
KeywordsHYDROLASE / ATPASE / NMH / DHU / ATP-BINDING / CREATINE METABOLISM
Function / homologyPHOSPHOAMINOPHOSPHONIC ACID-ADENYLATE ESTER / AMMONIUM ION
Function and homology information
Biological speciesGlutamicibacter protophormiae (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 3.13 Å
AuthorsStihle, M. / Benz, J. / Asztalos, P. / Rudolph, M.G.
Funding support Switzerland, 1items
OrganizationGrant numberCountry
F. Hoffmann-La Roche LTD Switzerland
CitationJournal: To be published
Title: Crystal Structure of a N-methylhydantoinase complex
Authors: Asztalos, P. / Meier, T. / Clairfeuille, T. / Rudolph, M.G.
History
DepositionOct 10, 2024Deposition site: RCSB / Processing site: RCSB
Revision 1.0Apr 15, 2026Provider: repository / Type: Initial release

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: N-methylhydantoinase
B: N-methylhydantoinase
C: N-methylhydantoinase
D: N-methylhydantoinase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)562,69116
Polymers560,4974
Non-polymers2,19412
Water28816
1
A: N-methylhydantoinase
B: N-methylhydantoinase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)281,3458
Polymers280,2482
Non-polymers1,0976
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
2
C: N-methylhydantoinase
D: N-methylhydantoinase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)281,3458
Polymers280,2482
Non-polymers1,0976
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Unit cell
Length a, b, c (Å)70.730, 84.250, 236.250
Angle α, β, γ (deg.)93.250, 94.820, 89.070
Int Tables number1
Space group name H-MP1
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails (eV)
11(chain A and (resid 1 through 690 or resid 693 through 1306))
21chain B
31(chain C and (resid 1 through 690 or resid 693 through 1306))
41(chain D and (resid 1 through 690 or resid 693 through 1306))

NCS domain segments:

Ens-ID: 1

Dom-IDComponent-IDBeg auth comp-IDBeg label comp-IDEnd auth comp-IDEnd label comp-IDSelection detailsAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
11METMETPROPRO(chain A and (resid 1 through 690 or resid 693 through 1306))AA1 - 6901 - 690
12THRTHRNH4NH4(chain A and (resid 1 through 690 or resid 693 through 1306))AA - G693 - 1303693
21METMETNH4NH4chain BBB - J1 - 13031
31METMETPROPRO(chain C and (resid 1 through 690 or resid 693 through 1306))CC1 - 6901 - 690
32THRTHRNH4NH4(chain C and (resid 1 through 690 or resid 693 through 1306))CC - M693 - 1303693
41METMETPROPRO(chain D and (resid 1 through 690 or resid 693 through 1306))DD1 - 6901 - 690
42THRTHRNH4NH4(chain D and (resid 1 through 690 or resid 693 through 1306))DD - P693 - 1303693

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Components

#1: Protein
N-methylhydantoinase


Mass: 140124.172 Da / Num. of mol.: 4
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Glutamicibacter protophormiae (bacteria)
Plasmid: pBP010 / Production host: Escherichia coli (E. coli) / Strain (production host): HB101
#2: Chemical
ChemComp-ANP / PHOSPHOAMINOPHOSPHONIC ACID-ADENYLATE ESTER


Mass: 506.196 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: C10H17N6O12P3 / Feature type: SUBJECT OF INVESTIGATION / Comment: AMP-PNP, energy-carrying molecule analogue*YM
#3: Chemical
ChemComp-MG / MAGNESIUM ION


Mass: 24.305 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: Mg / Feature type: SUBJECT OF INVESTIGATION
#4: Chemical
ChemComp-NH4 / AMMONIUM ION


Mass: 18.038 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: H4N
#5: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 16 / 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: 2.5 Å3/Da / Density % sol: 50.77 %
Crystal growTemperature: 293 K / Method: vapor diffusion, sitting drop / pH: 9
Details: 8 mg/ml protein in 20mM HEPES/NaOH pH 7.4, 0.1 M NaCl, 100 mM MgCl2, 30mM NH4Cl mixed 1+1 with 0.1 M Bicine/NaOH pH 9.0, 20 %w/v PEG 6000, 0.2 M MgCl2, 2 mM ADPNP

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: SLS / Beamline: X10SA / Wavelength: 0.99986 Å
DetectorType: PSI PILATUS 6M / Detector: PIXEL / Date: May 26, 2016
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.99986 Å / Relative weight: 1
ReflectionResolution: 3.13→49.38 Å / Num. obs: 92887 / % possible obs: 97.1 % / Redundancy: 1.8 % / Biso Wilson estimate: 54.12 Å2 / CC1/2: 0.926 / Rmerge(I) obs: 0.222 / Rrim(I) all: 0.314 / Χ2: 0.859 / Net I/σ(I): 3.58 / Num. measured all: 167171 / Scaling rejects: 37
Reflection shell

Diffraction-ID: 1

Resolution (Å)Redundancy (%)Rmerge(I) obsMean I/σ(I) obsNum. measured obsNum. possibleNum. unique obsCC1/2Rrim(I) all% possible all
3.13-3.211.670.9430.7511569715669270.3111.33496.8
3.21-3.31.7650.8780.8611651683466000.3221.24296.6
3.3-3.391.8580.6941.1312186676865570.4220.98196.9
3.39-3.51.8570.5351.4511671650462840.5480.75696.6
3.5-3.611.8460.4431.7411180626860550.6410.62796.6
3.61-3.741.8270.3672.0710850613159380.7130.51996.9
3.74-3.881.8280.3032.4810466588657250.7860.42897.3
3.88-4.041.7740.2392.969727567854830.8630.33896.6
4.04-4.221.6680.1953.548894546753320.8990.27597.5
4.22-4.431.7890.1584.419156523951190.9290.22397.7
4.43-4.671.8830.1395.199022491447920.9420.19697.5
4.67-4.951.8550.1355.228409465645320.9480.1997.3
4.95-5.291.8470.1444.857902439042790.9410.20497.5
5.29-5.711.8140.1564.527191408039640.9290.2297.2
5.71-6.261.6470.1643.996030375236610.9250.23397.6
6.26-71.8080.1384.956097344733720.9450.19697.8
7-8.081.890.0927.325421296228690.9760.1396.9
8.08-9.91.8240.05411.34509254624720.9910.07697.1
9.9-141.6910.04313.523261197219290.9930.0697.8
14-49.381.9850.03816.48197910539970.9950.05494.7

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Processing

Software
NameVersionClassificationNB
XSCALEdata scaling
PHENIX1.21.2_5419refinement
PDB_EXTRACT3.28data extraction
XDSdata reduction
PHASERphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: inhouse model

Resolution: 3.13→49.38 Å / SU ML: 0.52 / Cross valid method: THROUGHOUT / σ(F): 1.96 / Phase error: 28.27 / Stereochemistry target values: ML
Details: large conformational changes upon ADPNP binding; not all chains have density for an NH4+ ion near residue 959;
RfactorNum. reflection% reflectionSelection details
Rfree0.2645 4477 4.83 %RANDOM
Rwork0.223 88273 --
obs0.2249 92750 96.95 %-
Solvent computationShrinkage radii: 0.8 Å / VDW probe radii: 1 Å / Solvent model: FLAT BULK SOLVENT MODEL
Displacement parametersBiso max: 207.86 Å2 / Biso mean: 55.2702 Å2 / Biso min: 22.07 Å2
Refinement stepCycle: final / Resolution: 3.13→49.38 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms39140 0 132 16 39288
Biso mean--53.12 54.01 -
Num. residues----5118
Refine LS restraints NCS
Ens-IDDom-IDAuth asym-IDNumberRefine-IDRmsType
11A24356X-RAY DIFFRACTION5.357TORSIONAL
12B24356X-RAY DIFFRACTION5.357TORSIONAL
13C24356X-RAY DIFFRACTION5.357TORSIONAL
14D24356X-RAY DIFFRACTION5.357TORSIONAL
LS refinement shell

Refine-ID: X-RAY DIFFRACTION / Rfactor Rfree error: 0 / Total num. of bins used: 30

Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkNum. reflection all% reflection obs (%)
3.13-3.170.37471550.3592915307096
3.17-3.20.38991500.34852991314196
3.2-3.240.36171590.35092873303296
3.24-3.280.4231380.35392865300396
3.28-3.330.37641450.34352977312297
3.33-3.370.38991310.3273030316197
3.37-3.420.35831480.32022832298097
3.42-3.470.35831450.30572916306196
3.47-3.530.32781620.28622980314297
3.53-3.580.31971630.28062872303596
3.58-3.640.31361490.27312942309197
3.64-3.710.30511750.27092939311497
3.71-3.780.31241460.26612924307097
3.78-3.860.31631570.25972929308697
3.86-3.940.31341570.24712963312097
3.94-4.030.27971440.22812909305397
4.03-4.140.23771750.21342945312097
4.14-4.250.26521580.19762951310998
4.25-4.370.22181440.18572963310798
4.37-4.510.23541220.17592961308397
4.51-4.670.20161220.16343022314498
4.67-4.860.19181110.16952952306397
4.86-5.080.20881300.17193003313397
5.08-5.350.22851580.17882950310897
5.35-5.690.2231520.18632938309097
5.69-6.120.22421610.20432960312198
6.12-6.740.22571620.19452938310098
6.74-7.710.2311250.1663000312597
7.71-9.70.17081530.13532925307897
9.7-49.380.16851800.15272908308897
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)
10.16340.0463-0.02240.1874-0.10680.19220.09640.015-0.09080.1153-0.0962-0.048-0.00080.08420.06610.2170.0215-0.09480.1106-0.00930.208-11.781919.462149.4948
20.26180.01270.13020.4038-0.23190.5178-0.0426-0.0017-0.06740.0239-0.0306-0.08980.0405-0.0278-0.28140.18750.0391-0.02840.0962-0.0250.2096-27.33738.114338.4458
30.2672-0.04730.07550.04620.06450.2366-0.01110.1122-0.0050.01080.09460.1073-0.0124-0.06730.060.2133-0.0123-0.00320.12620.01920.1938-27.444644.378927.7688
40.0862-0.05330.27710.2517-0.130.4234-0.00990.39790.2746-0.03970.3070.2595-0.24930.05070.29640.2195-0.0727-0.02740.29020.17160.2054-15.715588.6425-32.771
50.3813-0.30940.12320.4130.19890.5867-0.0510.80060.40950.1995-0.1854-0.31650.1244-0.195-0.54780.1529-0.1166-0.22280.6550.20910.27716.23881.8822-40.8546
60.1398-0.19140.24080.4695-0.21990.3339-0.03830.44110.26840.11520.00060.42950.1074-0.0493-0.26780.0758-0.0369-0.17210.44160.20780.1862-19.65370.4254-39.7747
70.7476-0.0315-0.10730.18-0.25110.22540.02990.0707-0.03710.084-0.0186-0.0194-0.06360.06240.10750.151-0.05280.00550.1916-0.03390.1247-0.346569.6494-9.3058
80.1923-0.0125-0.2140.2944-0.07250.1198-0.0002-0.09940.0142-0.2715-0.02490.00190.0256-0.1195-0.29620.31260.0174-0.07550.1922-0.04920.1874-10.231347.356383.7003
90.14040.03870.12310.96650.45310.5108-0.0658-0.0603-0.083-0.0031-0.0622-0.07510.0492-0.0372-0.18830.22370.0028-0.00570.1728-0.02890.2664.476637.491996.7558
100.165-0.11440.07020.09490.1970.34130.0496-0.0367-0.0364-0.01840.0123-0.0662-0.07070.07710.32220.16760.0245-0.0070.2012-0.04490.17195.651673.0619105.2514
110.0647-0.06-0.0550.08320.03370.11770.0019-0.13640.09150.01820.1602-0.1598-0.18380.14790.09720.33990.0821-0.0520.3878-0.07690.0876-9.7934117.3903177.808
120.0286-0.01360.03460.0970.00230.15820.0179-0.09580.0877-0.04390.0523-0.0418-0.1311-0.0190.04820.3164-0.09610.06960.2195-0.06080.253-4.2348120.2365149.5117
130.148-0.1042-0.04140.13210.12130.52440.1452-0.18920.0390.06390.109-0.0193-0.3061-0.01780.33220.2014-0.036-0.010.30230.01270.1014-9.9792109.6499170.7419
140.03470.0928-0.11850.3099-0.33190.44660.0185-0.47230.3862-0.2771-0.18110.24340.1113-0.1879-0.17740.25420.0664-0.13630.4273-0.21140.4511-33.1268112.6405176.1158
150.21620.03470.25540.33470.17760.28820.1735-0.34270.1136-0.04630.0283-0.17020.14010.0062-0.00180.0545-0.045-0.07420.539-0.08690.2162-6.821499.4401174.9799
160.1882-0.02330.02010.03340.0420.1437-0.073-0.25-0.1985-0.0208-0.09020.06910.00310.1044-0.22380.06720.0189-0.07290.11630.1664-0.1567-18.593483.9988147.3568
170.13280.06430.09590.31040.15040.0750.0296-0.0397-0.1253-0.04830.06430.11730.0662-0.2160.03250.08810.0521-0.01130.37990.10950.1995-29.806391.062139.5401
180.02340.00950.04080.08410.05430.0666-0.0843-0.0149-0.1369-0.08050.0307-0.0218-0.0692-0.0544-0.00720.29670.06150.01330.31560.05260.2701-24.147198.261135.5833
190.0878-0.0001-0.06130.05550.01410.02370.0670.17070.104-0.16570.05520.0141-0.2572-0.09550.16580.35450.05760.06080.17950.07680.2124-18.0537117.0784134.5498
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection detailsAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1chain 'A' and (resid 1 through 264 )A1 - 264
2X-RAY DIFFRACTION2chain 'A' and (resid 265 through 703 )A265 - 703
3X-RAY DIFFRACTION3chain 'A' and (resid 704 through 1280 )A704 - 1280
4X-RAY DIFFRACTION4chain 'B' and (resid 1 through 291 )B1 - 291
5X-RAY DIFFRACTION5chain 'B' and (resid 292 through 478 )B292 - 478
6X-RAY DIFFRACTION6chain 'B' and (resid 479 through 649 )B479 - 649
7X-RAY DIFFRACTION7chain 'B' and (resid 650 through 1280 )B650 - 1280
8X-RAY DIFFRACTION8chain 'C' and (resid 1 through 264 )C1 - 264
9X-RAY DIFFRACTION9chain 'C' and (resid 265 through 744 )C265 - 744
10X-RAY DIFFRACTION10chain 'C' and (resid 745 through 1280 )C745 - 1280
11X-RAY DIFFRACTION11chain 'D' and (resid 1 through 95 )D1 - 95
12X-RAY DIFFRACTION12chain 'D' and (resid 96 through 202 )D96 - 202
13X-RAY DIFFRACTION13chain 'D' and (resid 203 through 332 )D203 - 332
14X-RAY DIFFRACTION14chain 'D' and (resid 333 through 478 )D333 - 478
15X-RAY DIFFRACTION15chain 'D' and (resid 479 through 675 )D479 - 675
16X-RAY DIFFRACTION16chain 'D' and (resid 676 through 870 )D676 - 870
17X-RAY DIFFRACTION17chain 'D' and (resid 871 through 997 )D871 - 997
18X-RAY DIFFRACTION18chain 'D' and (resid 998 through 1099 )D998 - 1099
19X-RAY DIFFRACTION19chain 'D' and (resid 1100 through 1280 )D0

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