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- PDB-5ejf: Crystal structure of NAD kinase P101A mutant from Listeria monocy... -

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Basic information

Entry
Database: PDB / ID: 5ejf
TitleCrystal structure of NAD kinase P101A mutant from Listeria monocytogenes
ComponentsNAD kinase 1
KeywordsTRANSFERASE / Gram-positive NAD Kinase / allostery / citrate
Function / homology
Function and homology information


NAD+ kinase / NADP biosynthetic process / NAD+ kinase activity / NAD metabolic process / NAD binding / phosphorylation / ATP binding / metal ion binding / cytoplasm
Similarity search - Function
Probable inorganic polyphosphate/atp-NAD kinase; domain 1 / Probable inorganic polyphosphate/atp-NAD kinase; domain 2 / ATP-NAD kinase C-terminal domain / NAD kinase / ATP-NAD kinase, PpnK-type, C-terminal / ATP-NAD kinase N-terminal domain / Inorganic polyphosphate/ATP-NAD kinase, N-terminal / NAD kinase/diacylglycerol kinase-like domain superfamily / Tumour Suppressor Smad4 / Sandwich ...Probable inorganic polyphosphate/atp-NAD kinase; domain 1 / Probable inorganic polyphosphate/atp-NAD kinase; domain 2 / ATP-NAD kinase C-terminal domain / NAD kinase / ATP-NAD kinase, PpnK-type, C-terminal / ATP-NAD kinase N-terminal domain / Inorganic polyphosphate/ATP-NAD kinase, N-terminal / NAD kinase/diacylglycerol kinase-like domain superfamily / Tumour Suppressor Smad4 / Sandwich / Rossmann fold / 3-Layer(aba) Sandwich / Mainly Beta / Alpha Beta
Similarity search - Domain/homology
Biological speciesListeria monocytogenes (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.12 Å
AuthorsPoncet-Montange, G. / Assairi, L. / Gelin, M. / Pochet, S. / Labesse, G.
Funding support France, 1items
OrganizationGrant numberCountry
French National Research Agency France
Citation
Journal: to be published
Title: Crystal structure of NAD kinase 1 from Listeria monocytogenes: P101A mutant
Authors: Poncet-Montange, G. / Assairi, L. / Gelin, M. / Pochet, S. / Labesse, G.
#1: Journal: To Be Published
Title: Molecular basis of NAD kinase catalysis reveals citrate is an allosteric regulator.
Authors: Poncet-Montange, G. / Assairi, L. / Gelin, M. / Pochet, S. / Labesse, G.
History
DepositionNov 1, 2015Deposition site: RCSB / Processing site: PDBE
Revision 1.0Nov 9, 2016Provider: repository / Type: Initial release
Revision 1.1Sep 6, 2017Group: Author supporting evidence / Data collection / Category: diffrn_radiation_wavelength / pdbx_audit_support / Item: _pdbx_audit_support.funding_organization
Revision 1.2Jan 10, 2024Group: 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: NAD kinase 1
B: NAD kinase 1
C: NAD kinase 1
D: NAD kinase 1


Theoretical massNumber of molelcules
Total (without water)124,0774
Polymers124,0774
Non-polymers00
Water9,152508
1


  • Idetical with deposited unit
  • defined by author&software
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area7450 Å2
ΔGint-46 kcal/mol
Surface area41040 Å2
MethodPISA
Unit cell
Length a, b, c (Å)67.042, 119.174, 67.593
Angle α, β, γ (deg.)90.000, 100.980, 90.000
Int Tables number4
Space group name H-MP1211
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11chain A
21chain B
31chain C
41chain D

NCS domain segments:

Component-ID: 1 / Ens-ID: 1 / Beg auth comp-ID: MET / Beg label comp-ID: MET

Dom-IDEnd auth comp-IDEnd label comp-IDSelection detailsAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
1ILEILEchain AAA1 - 2621 - 262
2HISHISchain BBB1 - 2671 - 267
3GLUGLUchain CCC1 - 2631 - 263
4GLUGLUchain DDD1 - 2631 - 263

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Components

#1: Protein
NAD kinase 1 / ATP-dependent NAD kinase


Mass: 31019.244 Da / Num. of mol.: 4 / Fragment: NAD Kinase 1-264
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Listeria monocytogenes (bacteria) / Gene: nadK1, lmo0968 / Plasmid: pLA15.3.3 / Production host: Escherichia coli BL21(DE3) (bacteria) / References: UniProt: Q8Y8D7, NAD+ kinase
#2: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 508 / 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: 2.14 Å3/Da / Density % sol: 42.43 %
Crystal growTemperature: 294 K / Method: vapor diffusion, hanging drop / pH: 5.1
Details: 50 mM Sodium Bromide, 150 mM tri-sodium citrate dihydrate, 14-16% w/v polyethylene glycol 400
PH range: 5.1-5.4

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: ESRF / Beamline: ID14-2 / Wavelength: 0.933 Å
DetectorType: ADSC QUANTUM 4 / Detector: CCD / Date: Sep 16, 2006
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.933 Å / Relative weight: 1
ReflectionResolution: 2.04→66.355 Å / Num. all: 58778 / Num. obs: 58778 / % possible obs: 99.6 % / Redundancy: 6 % / Rpim(I) all: 0.048 / Rrim(I) all: 0.119 / Rsym value: 0.109 / Net I/av σ(I): 4.77 / Net I/σ(I): 11.2 / Num. measured all: 351556
Reflection shell

Diffraction-ID: 1 / Rejects: _

Resolution (Å)Redundancy (%)Rmerge(I) obsMean I/σ(I) obsNum. measured allNum. unique allRpim(I) allRsym valueNet I/σ(I) obs% possible all
2.12-2.235.40.6331.24565584700.2940.6332.698.5
2.23-2.376.20.4631.65002281290.20.4633.9100
2.37-2.536.20.3192.34706176380.1370.3195.499.9
2.53-2.746.20.2023.64375470870.0870.202899.9
2.74-36.10.14654023265480.0630.14610.799.7
3-3.356.10.0957.13638359420.0410.09515.699.8
3.35-3.8760.0728.63142452200.0310.07220.699.8
3.87-4.745.90.0747.42632944380.0320.07423.899.6
4.74-6.75.60.0658.11915934010.0290.06523.898.9
6.7-34.7766.10.0527.81153719050.0230.05227.699.4

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Processing

Software
NameVersionClassification
PHENIXrefinement
MOSFLMdata reduction
SCALA3.3.21data scaling
PDB_EXTRACT3.15data extraction
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 2I1W
Resolution: 2.12→34.776 Å / SU ML: 0.31 / Cross valid method: FREE R-VALUE / σ(F): 0.03 / Phase error: 29.53 / Stereochemistry target values: ML
RfactorNum. reflection% reflection
Rfree0.2685 5887 5.08 %
Rwork0.2116 110074 -
obs0.2145 58737 99.44 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL
Displacement parametersBiso max: 135.39 Å2 / Biso mean: 45.4889 Å2 / Biso min: 9.62 Å2
Refinement stepCycle: final / Resolution: 2.12→34.776 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms7866 0 0 508 8374
Biso mean---44.28 -
Num. residues----1008
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.0118048
X-RAY DIFFRACTIONf_angle_d1.23910890
X-RAY DIFFRACTIONf_chiral_restr0.0651218
X-RAY DIFFRACTIONf_plane_restr0.0081384
X-RAY DIFFRACTIONf_dihedral_angle_d13.7512853
Refine LS restraints NCS
Ens-IDDom-IDAuth asym-IDNumberRefine-IDRmsType
11A4518X-RAY DIFFRACTION11.94TORSIONAL
12B4518X-RAY DIFFRACTION11.94TORSIONAL
13C4518X-RAY DIFFRACTION11.94TORSIONAL
14D4518X-RAY DIFFRACTION11.94TORSIONAL
LS refinement shell

Refine-ID: X-RAY DIFFRACTION / Total num. of bins used: 30

Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkNum. reflection all% reflection obs (%)
2.1197-2.14380.37041680.28163467363593
2.1438-2.1690.33522080.27833593380197
2.169-2.19540.34652010.282836493850100
2.1954-2.22320.33461760.275437253901100
2.2232-2.25240.38232170.270437343951100
2.2524-2.28330.33782080.265735693777100
2.2833-2.31590.32221780.244337973975100
2.3159-2.35050.30372280.23735733801100
2.3505-2.38720.31782050.252937063911100
2.3872-2.42630.34981780.249437103888100
2.4263-2.46810.35031660.260337053871100
2.4681-2.5130.33761970.242937143911100
2.513-2.56130.31161780.223236613839100
2.5613-2.61360.30522280.213236843912100
2.6136-2.67040.29981850.218336853870100
2.6704-2.73250.27342110.233136903901100
2.7325-2.80080.2952130.220436643877100
2.8008-2.87650.28651980.229536413839100
2.8765-2.96110.26092160.222336703886100
2.9611-3.05660.31721950.224637613956100
3.0566-3.16580.32831920.222336253817100
3.1658-3.29250.26651950.214336493844100
3.2925-3.44220.27141750.209837263901100
3.4422-3.62350.22552210.186236133834100
3.6235-3.85020.22161920.181537173909100
3.8502-4.14710.23851870.176736763863100
4.1471-4.56360.21741900.174337043894100
4.5636-5.2220.19372170.16993638385599
5.222-6.5720.25692050.20323619382499
6.572-34.78040.21721590.213137093868100
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)
10.01940.00950.01850.0036-0.00650.01950.12410.0815-0.09530.02790.04-0.05810.03080.0001-0.00710.5148-0.12260.12010.08120.02590.3682.3076-40.4126-28.8538
20.06120.0060.0355-0-0.01090.01380.20280.0187-0.01920.0583-0.03080.08840.0692-0.0450.03870.2823-0.06040.07230.12760.00230.30441.1079-28.8503-26.1913
30.00620.0021-0.00570.0054-0.0114-0.00250.0880.0192-0.02780.0670.06470.04630.1122-0.01450.00010.30630.01320.00910.2054-0.00040.299313.1251-32.3243-31.3264
40.28430.17130.00860.11960.00670.13860.10970.0159-0.00650.0069-0.07590.12630.0191-0.04090.01390.15110.01690.01060.1141-0.01220.149811.6506-10.6748-37.5773
50.03330.00060.0140.0056-0.02350.02490.11120.02520.09370.0530.05990.145-0.0024-0.05250.02590.07480.0278-0.04490.12230.00850.27671.3747-8.5425-41.7274
60.05720.02920.06610.00680.02040.08560.29450.1485-0.11240.0512-0.0019-0.10030.06040.20660.07570.1717-0.24690.0932-0.21490.09470.203418.6246-24.9024-29.4329
70.00390.00370.00020.0009-0.00370.0009-0.03320.02670.143-0.0143-0.0432-0.0369-0.03810.0120.00010.5196-0.165-0.01930.4061-0.15660.522740.129120.0148-24.4487
80.00470.0002-0.01320.01270.02440.0602-0.1248-0.18470.1601-0.01430.08770.01510.01050.08590.00040.3604-0.2094-0.0140.3256-0.17850.264139.4779.1222-20.7648
90.0028-0.00050.00150.00370.00250.00380.0775-0.04320.1420.03090.07650.0235-0.01670.012700.3921-0.0768-0.02810.2037-0.03430.260530.952212.1731-29.5687
100.11020.03510.00130.03130.00570.09880.1165-0.14730.0650.08610.016-0.02470.11480.02470.07620.1522-0.015-0.05220.1342-0.02480.172834.01364.0843-22.7441
110.02060.03270.03340.01360.01320.03170.0670.00470.03790.0744-0.0089-0.08860.01430.05440.00160.12540.0106-0.02920.13260.00080.139734.4692-13.8318-37.9763
120.05740.0859-0.00730.08840.02170.02460.2747-0.0107-0.06360.1549-0.1593-0.1230.02540.09070.17840.16350.0116-0.05940.2013-0.00230.116739.7442-11.7306-29.2041
13-0.00940.00920.0144-0.00620.00420.00350.0482-0.0270.0879-0.0566-0.08110.08460.04930.0068-00.15130.00480.03760.1271-0.02520.279721.301210.0827-35.972
140.0256-0.01140.0030.0237-0.00930.00870.08460.11560.20960.05930.04170.07480.0069-0.03570.00020.33460.164-0.01160.38760.03370.38569.62326.9881-79.5459
150.1309-0.0816-0.01880.14180.01550.17960.11150.1598-0.15150.0092-0.14330.0747-0.0456-0.0630.04850.15280.0186-0.0160.2054-0.08610.153418.8501-18.3459-64.0649
160.0434-0.00420.00220.0222-0.0442-0.010.14280.1945-0.0372-0.0874-0.2487-0.0334-0.1-0.1099-0.00060.2410.0641-0.01890.3225-0.04280.215316.4014-6.879-69.1111
170.0092-0.00020.0013-0.00110.00130.0066-0.10270.1009-0.07940.0399-0.0185-0.02270.02420.02850.00020.49650.17940.00940.486-0.1440.575359.8596-31.2139-68.8887
180.0464-0.02290.02620.0161-0.01180.0180.02830.0002-0.01130.0570.1081-0.03450.02110.10920.0199-0.01350.24910.11750.4645-0.26460.48455.6994-20.9833-72.2119
190.00170.00070.00670.0014-0.00050.00470.0858-0.0185-0.05960.04270.07380.0442-0.0392-0.0709-0.00080.48620.02440.09750.4555-0.19170.461348.2305-32.4518-70.6499
20-0.001-0.0027-0.0145-0.00420.02920.00220.1850.11540.1131-0.0884-0.0339-0.06220.04550.011100.2220.01070.05870.3447-0.00630.212748.4334-6.4558-67.8338
210.0458-0.03960.0080.0239-0.02020.03630.02710.06290.0474-0.0395-0.0298-0.05530.00130.00600.17640.0070.03220.20260.02450.13437.3077-1.1689-57.47
220.0383-0.01770.01810.01840.01120.0913-0.00290.2882-0.02150.0595-0.03-0.1063-0.04260.1517-0.00960.172-0.01610.0180.3489-0.0270.227648.1176-6.4273-63.3451
230.0169-0.0036-0.0018-0.0011-0.0021-0.0013-0.01990.0179-0.0409-0.1077-0.0023-0.06070.06990.00250.00080.28650.047-0.04510.27440.00210.266333.1844-26.6955-61.5526
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection detailsAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1chain 'A' and (resid 1 through 26 )A0
2X-RAY DIFFRACTION2chain 'A' and (resid 27 through 61 )A0
3X-RAY DIFFRACTION3chain 'A' and (resid 62 through 79 )A0
4X-RAY DIFFRACTION4chain 'A' and (resid 80 through 216 )A0
5X-RAY DIFFRACTION5chain 'A' and (resid 217 through 238 )A0
6X-RAY DIFFRACTION6chain 'A' and (resid 239 through 262 )A0
7X-RAY DIFFRACTION7chain 'B' and (resid 1 through 22 )B0
8X-RAY DIFFRACTION8chain 'B' and (resid 23 through 61 )B0
9X-RAY DIFFRACTION9chain 'B' and (resid 62 through 79 )B0
10X-RAY DIFFRACTION10chain 'B' and (resid 80 through 120 )B0
11X-RAY DIFFRACTION11chain 'B' and (resid 121 through 198 )B0
12X-RAY DIFFRACTION12chain 'B' and (resid 199 through 247 )B0
13X-RAY DIFFRACTION13chain 'B' and (resid 248 through 267 )B0
14X-RAY DIFFRACTION14chain 'C' and (resid 1 through 100 )C0
15X-RAY DIFFRACTION15chain 'C' and (resid 101 through 216 )C0
16X-RAY DIFFRACTION16chain 'C' and (resid 217 through 263 )C0
17X-RAY DIFFRACTION17chain 'D' and (resid 1 through 26 )D0
18X-RAY DIFFRACTION18chain 'D' and (resid 27 through 68 )D0
19X-RAY DIFFRACTION19chain 'D' and (resid 69 through 91 )D0
20X-RAY DIFFRACTION20chain 'D' and (resid 92 through 122 )D0
21X-RAY DIFFRACTION21chain 'D' and (resid 123 through 216 )D0
22X-RAY DIFFRACTION22chain 'D' and (resid 217 through 252 )D0
23X-RAY DIFFRACTION23chain 'D' and (resid 253 through 263 )D0

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