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- PDB-1zmx: Crystal structure of D. melanogaster deoxyribonucleoside kinase N... -

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Basic information

Entry
Database: PDB / ID: 1zmx
TitleCrystal structure of D. melanogaster deoxyribonucleoside kinase N64D mutant in complex with thymidine
ComponentsDeoxynucleoside kinase
KeywordsTRANSFERASE / Drosophila melanogaster / dnk / N64D mutant
Function / homology
Function and homology information


deoxynucleoside kinase / Pyrimidine salvage / deoxynucleoside kinase activity / nucleoside salvage / uridine kinase activity / deoxycytidine kinase activity / nucleoside phosphate biosynthetic process / thymidine kinase activity / deoxyguanosine kinase activity / deoxyadenosine kinase activity ...deoxynucleoside kinase / Pyrimidine salvage / deoxynucleoside kinase activity / nucleoside salvage / uridine kinase activity / deoxycytidine kinase activity / nucleoside phosphate biosynthetic process / thymidine kinase activity / deoxyguanosine kinase activity / deoxyadenosine kinase activity / cytidine kinase activity / DNA biosynthetic process / kinase activity / phosphorylation / mitochondrion / ATP binding / cytoplasm
Similarity search - Function
Deoxynucleoside kinase / Deoxynucleoside kinase domain / Deoxynucleoside kinase / P-loop containing nucleotide triphosphate hydrolases / P-loop containing nucleoside triphosphate hydrolase / Rossmann fold / 3-Layer(aba) Sandwich / Alpha Beta
Similarity search - Domain/homology
THYMIDINE / Deoxynucleoside kinase
Similarity search - Component
Biological speciesDrosophila melanogaster (fruit fly)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 3.1 Å
AuthorsWelin, M. / Skovgaard, T. / Knecht, W. / Berenstein, D. / Munch-Petersen, B. / Piskur, J. / Eklund, H.
CitationJournal: Febs J. / Year: 2005
Title: Structural basis for the changed substrate specificity of Drosophila melanogaster deoxyribonucleoside kinase mutant N64D.
Authors: Welin, M. / Skovgaard, T. / Knecht, W. / Zhu, C. / Berenstein, D. / Munch-Petersen, B. / Piskur, J. / Eklund, H.
History
DepositionMay 11, 2005Deposition site: RCSB / Processing site: RCSB
Revision 1.0May 24, 2005Provider: repository / Type: Initial release
Revision 1.1Apr 30, 2008Group: Version format compliance
Revision 1.2Jul 13, 2011Group: Version format compliance
Revision 1.3Oct 20, 2021Group: Data collection / Database references ...Data collection / Database references / Derived calculations / Experimental preparation
Category: chem_comp / database_2 ...chem_comp / database_2 / exptl_crystal_grow / struct_ref_seq_dif / struct_site
Item: _chem_comp.type / _database_2.pdbx_DOI ..._chem_comp.type / _database_2.pdbx_DOI / _database_2.pdbx_database_accession / _exptl_crystal_grow.method / _struct_ref_seq_dif.details / _struct_site.pdbx_auth_asym_id / _struct_site.pdbx_auth_comp_id / _struct_site.pdbx_auth_seq_id
Revision 1.4Aug 23, 2023Group: Data collection / Refinement description
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / pdbx_initial_refinement_model / struct_ncs_dom_lim
Item: _struct_ncs_dom_lim.beg_auth_comp_id / _struct_ncs_dom_lim.end_auth_comp_id

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

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Assembly

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


Theoretical massNumber of molelcules
Total (without water)217,96824
Polymers215,2628
Non-polymers2,70616
Water0
1
A: Deoxynucleoside kinase
B: Deoxynucleoside kinase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)54,4926
Polymers53,8152
Non-polymers6774
Water0
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area4260 Å2
ΔGint-59 kcal/mol
Surface area17470 Å2
MethodPISA
2
C: Deoxynucleoside kinase
D: Deoxynucleoside kinase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)54,4926
Polymers53,8152
Non-polymers6774
Water0
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area4290 Å2
ΔGint-59 kcal/mol
Surface area17550 Å2
MethodPISA
3
E: Deoxynucleoside kinase
F: Deoxynucleoside kinase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)54,4926
Polymers53,8152
Non-polymers6774
Water0
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area4270 Å2
ΔGint-58 kcal/mol
Surface area17580 Å2
MethodPISA
4
G: Deoxynucleoside kinase
H: Deoxynucleoside kinase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)54,4926
Polymers53,8152
Non-polymers6774
Water0
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area4270 Å2
ΔGint-58 kcal/mol
Surface area17570 Å2
MethodPISA
Unit cell
Length a, b, c (Å)69.712, 70.340, 224.528
Angle α, β, γ (deg.)90.00, 90.69, 90.00
Int Tables number4
Space group name H-MP1211
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11A
21B
31C
41D
51E
61F
71G
81H
91A
101B
111C
121D
131E
141F
151G
161H
171A
181B
191C
201D
211E
221F
231G
241H
251A
261B
271C
281D
291E
301F
311G
321H
331A
341B
351C
361D
371E
381F
391G
401H
411A
421B
431C
441D
451E
461F
471G
481H
491A
501B
511C
521D
531E
541F
551G
561H
571A
581B
591C
601D
611E
621F
631G
641H
651A
661B
671C
681D
691E
701F
711G
721H
731A
741B
751C
761D
771E
781F
791G
801H
811A
821B
831C
841D
851E
861F
871G
881H
891A
901B
911C
921D
931E
941F
951G
961H
971A
981B
991C
1001D
1011E
1021F
1031G
1041H
1051A
1061B
1071C
1081D
1091E
1101F
1111G
1121H

NCS domain segments:

Ens-ID: 1

Dom-IDComponent-IDBeg auth comp-IDBeg label comp-IDEnd auth comp-IDEnd label comp-IDRefine codeAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
11THRTHRVALVAL1AA12 - 5412 - 54
21THRTHRVALVAL1BB12 - 5412 - 54
31THRTHRVALVAL1CC12 - 5412 - 54
41THRTHRVALVAL1DD12 - 5412 - 54
51THRTHRVALVAL1EE12 - 5412 - 54
61THRTHRVALVAL1FF12 - 5412 - 54
71THRTHRVALVAL1GG12 - 5412 - 54
81THRTHRVALVAL1HH12 - 5412 - 54
92GLUGLUGLUGLU3AA5555
102GLUGLUGLUGLU3BB5555
112GLUGLUGLUGLU3CC5555
122GLUGLUGLUGLU3DD5555
132GLUGLUGLUGLU3EE5555
142GLUGLUGLUGLU3FF5555
152GLUGLUGLUGLU3GG5555
162GLUGLUGLUGLU3HH5555
173LYSLYSLEULEU1AA56 - 6656 - 66
183LYSLYSLEULEU1BB56 - 6656 - 66
193LYSLYSLEULEU1CC56 - 6656 - 66
203LYSLYSLEULEU1DD56 - 6656 - 66
213LYSLYSLEULEU1EE56 - 6656 - 66
223LYSLYSLEULEU1FF56 - 6656 - 66
233LYSLYSLEULEU1GG56 - 6656 - 66
243LYSLYSLEULEU1HH56 - 6656 - 66
254GLUGLUGLUGLU3AA6767
264GLUGLUGLUGLU3BB6767
274GLUGLUGLUGLU3CC6767
284GLUGLUGLUGLU3DD6767
294GLUGLUGLUGLU3EE6767
304GLUGLUGLUGLU3FF6767
314GLUGLUGLUGLU3GG6767
324GLUGLUGLUGLU3HH6767
335LEULEULEULEU1AA68 - 12468 - 124
345LEULEULEULEU1BB68 - 12468 - 124
355LEULEULEULEU1CC68 - 12468 - 124
365LEULEULEULEU1DD68 - 12468 - 124
375LEULEULEULEU1EE68 - 12468 - 124
385LEULEULEULEU1FF68 - 12468 - 124
395LEULEULEULEU1GG68 - 12468 - 124
405LEULEULEULEU1HH68 - 12468 - 124
416GLUGLUGLNGLN3AA125 - 126125 - 126
426GLUGLUGLNGLN3BB125 - 126125 - 126
436GLUGLUGLNGLN3CC125 - 126125 - 126
446GLUGLUGLNGLN3DD125 - 126125 - 126
456GLUGLUGLNGLN3EE125 - 126125 - 126
466GLUGLUGLNGLN3FF125 - 126125 - 126
476GLUGLUGLNGLN3GG125 - 126125 - 126
486GLUGLUGLNGLN3HH125 - 126125 - 126
497GLYGLYILEILE1AA127 - 139127 - 139
507GLYGLYILEILE1BB127 - 139127 - 139
517GLYGLYILEILE1CC127 - 139127 - 139
527GLYGLYILEILE1DD127 - 139127 - 139
537GLYGLYILEILE1EE127 - 139127 - 139
547GLYGLYILEILE1FF127 - 139127 - 139
557GLYGLYILEILE1GG127 - 139127 - 139
567GLYGLYILEILE1HH127 - 139127 - 139
578GLUGLUGLUGLU3AA140 - 141140 - 141
588GLUGLUGLUGLU3BB140 - 141140 - 141
598GLUGLUGLUGLU3CC140 - 141140 - 141
608GLUGLUGLUGLU3DD140 - 141140 - 141
618GLUGLUGLUGLU3EE140 - 141140 - 141
628GLUGLUGLUGLU3FF140 - 141140 - 141
638GLUGLUGLUGLU3GG140 - 141140 - 141
648GLUGLUGLUGLU3HH140 - 141140 - 141
659SERSERALAALA1AA142 - 160142 - 160
669SERSERALAALA1BB142 - 160142 - 160
679SERSERALAALA1CC142 - 160142 - 160
689SERSERALAALA1DD142 - 160142 - 160
699SERSERALAALA1EE142 - 160142 - 160
709SERSERALAALA1FF142 - 160142 - 160
719SERSERALAALA1GG142 - 160142 - 160
729SERSERALAALA1HH142 - 160142 - 160
7310TYRTYRGLUGLU3AA161 - 162161 - 162
7410TYRTYRGLUGLU3BB161 - 162161 - 162
7510TYRTYRGLUGLU3CC161 - 162161 - 162
7610TYRTYRGLUGLU3DD161 - 162161 - 162
7710TYRTYRGLUGLU3EE161 - 162161 - 162
7810TYRTYRGLUGLU3FF161 - 162161 - 162
7910TYRTYRGLUGLU3GG161 - 162161 - 162
8010TYRTYRGLUGLU3HH161 - 162161 - 162
8111ARGARGILEILE1AA163 - 164163 - 164
8211ARGARGILEILE1BB163 - 164163 - 164
8311ARGARGILEILE1CC163 - 164163 - 164
8411ARGARGILEILE1DD163 - 164163 - 164
8511ARGARGILEILE1EE163 - 164163 - 164
8611ARGARGILEILE1FF163 - 164163 - 164
8711ARGARGILEILE1GG163 - 164163 - 164
8811ARGARGILEILE1HH163 - 164163 - 164
8912ARGARGPROPRO2AA165 - 176165 - 176
9012ARGARGPROPRO2BB165 - 176165 - 176
9112ARGARGPROPRO2CC165 - 176165 - 176
9212ARGARGPROPRO2DD165 - 176165 - 176
9312ARGARGPROPRO2EE165 - 176165 - 176
9412ARGARGPROPRO2FF165 - 176165 - 176
9512ARGARGPROPRO2GG165 - 176165 - 176
9612ARGARGPROPRO2HH165 - 176165 - 176
9713LEULEUGLNGLN1AA177 - 194177 - 194
9813LEULEUGLNGLN1BB177 - 194177 - 194
9913LEULEUGLNGLN1CC177 - 194177 - 194
10013LEULEUGLNGLN1DD177 - 194177 - 194
10113LEULEUGLNGLN1EE177 - 194177 - 194
10213LEULEUGLNGLN1FF177 - 194177 - 194
10313LEULEUGLNGLN1GG177 - 194177 - 194
10413LEULEUGLNGLN1HH177 - 194177 - 194
10514CYSCYSASPASP1AA200 - 206200 - 206
10614CYSCYSASPASP1BB200 - 206200 - 206
10714CYSCYSASPASP1CC200 - 206200 - 206
10814CYSCYSASPASP1DD200 - 206200 - 206
10914CYSCYSASPASP1EE200 - 206200 - 206
11014CYSCYSASPASP1FF200 - 206200 - 206
11114CYSCYSASPASP1GG200 - 206200 - 206
11214CYSCYSASPASP1HH200 - 206200 - 206

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Components

#1: Protein
Deoxynucleoside kinase / / Deoxyribonucleoside kinase / Dm-dNK / Multispecific deoxynucleoside kinase


Mass: 26907.691 Da / Num. of mol.: 8 / Fragment: residues 1-230 / Mutation: N64D
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Drosophila melanogaster (fruit fly) / Gene: dnk / Plasmid: pGEX-2T / Species (production host): Escherichia coli / Production host: Escherichia coli BL21 (bacteria) / Strain (production host): BL21 / References: UniProt: Q9XZT6, deoxynucleoside kinase
#2: Chemical
ChemComp-SO4 / SULFATE ION / Sulfate


Mass: 96.063 Da / Num. of mol.: 8 / Source method: obtained synthetically / Formula: SO4
#3: Chemical
ChemComp-THM / THYMIDINE / DEOXYTHYMIDINE / 2'-DEOXYTHYMIDINE / Thymidine


Type: DNA OH 5 prime terminus / Mass: 242.229 Da / Num. of mol.: 8 / Source method: obtained synthetically / Formula: C10H14N2O5

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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: 2.8 Å3/Da / Density % sol: 55 %
Crystal growTemperature: 288 K / Method: counter-diffusion / pH: 6.5
Details: MES, lithium sulphate, mPEG 2000, pH 6.5, Counter diffusion, temperature 288K

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: ESRF / Beamline: ID14-4 / Wavelength: 0.939 Å
DetectorType: ADSC QUANTUM 4 / Detector: CCD / Date: Sep 14, 2003 / Details: Mirrors
RadiationMonochromator: Mirrors / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.939 Å / Relative weight: 1
ReflectionResolution: 3.1→41.17 Å / Num. obs: 39975 / % possible obs: 99.9 % / Observed criterion σ(F): 1 / Observed criterion σ(I): 1
Reflection shellResolution: 3.1→3.27 Å / Redundancy: 3.7 % / Mean I/σ(I) obs: 3.4 / Num. unique all: 39562 / Rsym value: 0.38 / % possible all: 99.9

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Processing

Software
NameVersionClassification
REFMAC5.1.24refinement
MOSFLMdata reduction
CCP4(SCALA)data scaling
MOLREPphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: PDB entry 1OT3
Resolution: 3.1→41.17 Å / Cor.coef. Fo:Fc: 0.871 / Cor.coef. Fo:Fc free: 0.842 / SU B: 22.712 / SU ML: 0.399 / Cross valid method: THROUGHOUT / σ(F): 1 / ESU R Free: 0.526 / Stereochemistry target values: MAXIMUM LIKELIHOOD
RfactorNum. reflection% reflectionSelection details
Rfree0.2884 2005 5 %RANDOM
Rwork0.26858 ---
all0.2696 ---
obs0.26958 37947 99.96 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.4 Å / Solvent model: BABINET MODEL WITH MASK
Displacement parametersBiso mean: 37.441 Å2
Baniso -1Baniso -2Baniso -3
1--0.9 Å20 Å22.48 Å2
2--2.32 Å20 Å2
3----1.36 Å2
Refinement stepCycle: LAST / Resolution: 3.1→41.17 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms12536 0 176 0 12712
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0090.02113000
X-RAY DIFFRACTIONr_angle_refined_deg1.1341.96617582
X-RAY DIFFRACTIONr_dihedral_angle_1_deg5.31451482
X-RAY DIFFRACTIONr_chiral_restr0.090.21910
X-RAY DIFFRACTIONr_gen_planes_refined0.0040.029688
X-RAY DIFFRACTIONr_nbd_refined0.2020.25468
X-RAY DIFFRACTIONr_xyhbond_nbd_refined0.1340.2304
X-RAY DIFFRACTIONr_symmetry_vdw_refined0.3130.272
X-RAY DIFFRACTIONr_symmetry_hbond_refined0.4580.22
X-RAY DIFFRACTIONr_mcbond_it0.6351.57494
X-RAY DIFFRACTIONr_mcangle_it1.212212184
X-RAY DIFFRACTIONr_scbond_it1.24735506
X-RAY DIFFRACTIONr_scangle_it2.2544.55398
Refine LS restraints NCS

Ens-ID: 1 / Refine-ID: X-RAY DIFFRACTION

Dom-IDAuth asym-IDNumberTypeRms dev position (Å)Weight position
1A1463tight positional0.010.01
2B1463tight positional0.010.01
3C1463tight positional0.010.01
4D1463tight positional0.010.01
5E1463tight positional0.040.01
6F1463tight positional0.010.01
7G1463tight positional0.010.01
8H1463tight positional0.010.01
1A25medium positional0.880.5
2B25medium positional1.140.5
3C25medium positional1.120.5
4D25medium positional0.970.5
5E25medium positional1.910.5
6F25medium positional0.910.5
7G25medium positional1.350.5
8H25medium positional1.090.5
1A43loose positional0.755
2B43loose positional1.055
3C43loose positional1.45
4D43loose positional1.255
5E43loose positional1.055
6F43loose positional1.075
7G43loose positional0.955
8H43loose positional0.965
1A1463tight thermal0.120.5
2B1463tight thermal0.080.5
3C1463tight thermal0.10.5
4D1463tight thermal0.070.5
5E1463tight thermal0.080.5
6F1463tight thermal0.070.5
7G1463tight thermal0.090.5
8H1463tight thermal0.080.5
1A25medium thermal0.32
2B25medium thermal0.612
3C25medium thermal0.482
4D25medium thermal0.342
5E25medium thermal0.652
6F25medium thermal0.462
7G25medium thermal0.472
8H25medium thermal0.982
1A43loose thermal2.4710
2B43loose thermal2.8410
3C43loose thermal1.6410
4D43loose thermal0.8510
5E43loose thermal2.8310
6F43loose thermal1.8310
7G43loose thermal1.6910
8H43loose thermal1.510
LS refinement shellResolution: 3.1→3.18 Å / Total num. of bins used: 20 /
RfactorNum. reflection
Rfree0.302 158
Rwork0.311 2794

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