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- PDB-9k2n: Crystal structure of Glutamine Synthetase-apo -

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Basic information

Entry
Database: PDB / ID: 9k2n
TitleCrystal structure of Glutamine Synthetase-apo
ComponentsGlutamine synthetase
KeywordsLIGASE / Acinetobacter baumannii / Glutamine synthetase / Biosynthetic protein
Function / homology
Function and homology information


nitrogen utilization / glutamine synthetase / glutamine biosynthetic process / glutamine synthetase activity / ATP binding / membrane / cytoplasm
Similarity search - Function
Glutamine synthetase class-I, adenylation site / Glutamine synthetase class-I adenylation site. / Glutamine synthetase type I / Glutamine synthetase, N-terminal conserved site / Glutamine synthetase signature 1. / Glutamine synthetase, beta-Grasp domain / Glutamine synthetase, glycine-rich site / Glutamine synthetase putative ATP-binding region signature. / Glutamine synthetase (GS) beta-grasp domain profile. / Glutamine synthetase, N-terminal domain superfamily ...Glutamine synthetase class-I, adenylation site / Glutamine synthetase class-I adenylation site. / Glutamine synthetase type I / Glutamine synthetase, N-terminal conserved site / Glutamine synthetase signature 1. / Glutamine synthetase, beta-Grasp domain / Glutamine synthetase, glycine-rich site / Glutamine synthetase putative ATP-binding region signature. / Glutamine synthetase (GS) beta-grasp domain profile. / Glutamine synthetase, N-terminal domain superfamily / Glutamine synthetase, catalytic domain / Glutamine synthetase, N-terminal domain / Glutamine synthetase, catalytic domain / Glutamine synthetase (GS) catalytic domain profile. / Glutamine synthetase, catalytic domain / Glutamine synthetase/guanido kinase, catalytic domain
Similarity search - Domain/homology
TRIETHYLENE GLYCOL / Glutamine synthetase
Similarity search - Component
Biological speciesAcinetobacter baumannii AB0057 (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 3.4 Å
AuthorsChandrasekaran, P. / Killivalavan, A. / Vaigundan, D. / Yuvaraj, I. / Manju, B. / Sekar, K.
Funding support India, 1items
OrganizationGrant numberCountry
Department of Science & Technology (DST, India)Meit/R&D/HPC/2(1)/2014 India
CitationJournal: To Be Published
Title: Structure of glutamine synthetase from ESKAPE pathogen Acinetobacter baumannii
Authors: Chandrasekaran, P. / Killivalavan, A. / Vaigundan, D. / Yuvaraj, I. / Manju, B. / Sekar, K.
History
DepositionOct 17, 2024Deposition site: PDBJ / Processing site: PDBJ
Revision 1.0Mar 19, 2025Provider: repository / Type: Initial release

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Glutamine synthetase
B: Glutamine synthetase
C: Glutamine synthetase
D: Glutamine synthetase
E: Glutamine synthetase
F: Glutamine synthetase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)313,93327
Polymers312,7846
Non-polymers1,14921
Water00
1
A: Glutamine synthetase
B: Glutamine synthetase
C: Glutamine synthetase
D: Glutamine synthetase
E: Glutamine synthetase
F: Glutamine synthetase
hetero molecules

A: Glutamine synthetase
B: Glutamine synthetase
C: Glutamine synthetase
D: Glutamine synthetase
E: Glutamine synthetase
F: Glutamine synthetase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)627,86554
Polymers625,56812
Non-polymers2,29742
Water0
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
crystal symmetry operation8_554-y,-x,-z-1/21
Buried area82900 Å2
ΔGint-644 kcal/mol
Surface area173520 Å2
MethodPISA
Unit cell
Length a, b, c (Å)260.909, 260.909, 154.18
Angle α, β, γ (deg.)90, 90, 90
Int Tables number96
Space group name H-MP43212
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11A
21B
32A
42C
53A
63D
74A
84E
95A
105F
116B
126C
137B
147D
158B
168E
179B
189F
1910C
2010D
2111C
2211E
2312C
2412F
2513D
2613E
2714D
2814F
2915E
3015F

NCS domain segments:

Beg auth comp-ID: ALA / Beg label comp-ID: ALA / End auth comp-ID: LEU / End label comp-ID: LEU / Auth seq-ID: 2 - 469 / Label seq-ID: 1 - 468

Dom-IDComponent-IDEns-IDAuth asym-IDLabel asym-ID
111AA
211BB
322AA
422CC
533AA
633DD
744AA
844EE
955AA
1055FF
1166BB
1266CC
1377BB
1477DD
1588BB
1688EE
1799BB
1899FF
191010CC
201010DD
211111CC
221111EE
231212CC
241212FF
251313DD
261313EE
271414DD
281414FF
291515EE
301515FF

NCS ensembles :
IDDetails
1Local NCS retraints between domains: 1 2
2Local NCS retraints between domains: 3 4
3Local NCS retraints between domains: 5 6
4Local NCS retraints between domains: 7 8
5Local NCS retraints between domains: 9 10
6Local NCS retraints between domains: 11 12
7Local NCS retraints between domains: 13 14
8Local NCS retraints between domains: 15 16
9Local NCS retraints between domains: 17 18
10Local NCS retraints between domains: 19 20
11Local NCS retraints between domains: 21 22
12Local NCS retraints between domains: 23 24
13Local NCS retraints between domains: 25 26
14Local NCS retraints between domains: 27 28
15Local NCS retraints between domains: 29 30

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Components

#1: Protein
Glutamine synthetase


Mass: 52130.668 Da / Num. of mol.: 6
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Acinetobacter baumannii AB0057 (bacteria)
Gene: AB57_2783 / Plasmid: pET28a+ / Production host: Escherichia coli BL21(DE3) (bacteria) / References: UniProt: A0A7U3Y893, glutamine synthetase
#2: Chemical ChemComp-PGE / TRIETHYLENE GLYCOL


Mass: 150.173 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C6H14O4 / Feature type: SUBJECT OF INVESTIGATION
#3: Chemical
ChemComp-GOL / GLYCEROL / GLYCERIN / PROPANE-1,2,3-TRIOL


Mass: 92.094 Da / Num. of mol.: 7 / Source method: obtained synthetically / Formula: C3H8O3 / Feature type: SUBJECT OF INVESTIGATION
#4: Chemical
ChemComp-MG / MAGNESIUM ION


Mass: 24.305 Da / Num. of mol.: 12 / Source method: isolated from a natural source / Formula: Mg / Feature type: SUBJECT OF INVESTIGATION
#5: Chemical ChemComp-EDO / 1,2-ETHANEDIOL / ETHYLENE GLYCOL


Mass: 62.068 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C2H6O2 / Feature type: SUBJECT OF INVESTIGATION
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: 4.06 Å3/Da / Density % sol: 71.04 % / Description: Cuboid structure
Crystal growTemperature: 293 K / Method: vapor diffusion, hanging drop / pH: 9
Details: 0.1M NaCl, 0.1M BICINE pH 9.0, 5mM MgCl2, 20% v/v polyethylene glycol monomethyl ether 550
PH range: 8.5 - 9.3

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Data collection

DiffractionMean temperature: 100 K / Serial crystal experiment: N
Diffraction sourceSource: SYNCHROTRON / Site: ELETTRA / Beamline: 11.2C / Wavelength: 1 Å
DetectorType: DECTRIS PILATUS 6M / Detector: PIXEL / Date: Apr 24, 2024
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 1 Å / Relative weight: 1
ReflectionResolution: 3.4→99.59 Å / Num. obs: 72108 / % possible obs: 98.5 % / Redundancy: 6.8 % / CC1/2: 0.967 / Net I/σ(I): 9
Reflection shellResolution: 3.4→3.47 Å / Redundancy: 7.1 % / Mean I/σ(I) obs: 2.6 / Num. unique obs: 4374 / CC1/2: 0.36 / % possible all: 98.1

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Processing

Software
NameVersionClassification
REFMAC5.8.0430 (refmacat 0.4.100)refinement
autoPX9.0.001data processing
iMOSFLMccp4 9.0.001data reduction
Aimlessccp4 9.0.001data scaling
MOLREPccp4 9.0.001phasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT / Resolution: 3.4→93.215 Å / Cor.coef. Fo:Fc: 0.904 / Cor.coef. Fo:Fc free: 0.881 / SU B: 77.444 / SU ML: 0.533 / Cross valid method: THROUGHOUT / ESU R Free: 0.532
Details: Hydrogens have been added in their riding positions
RfactorNum. reflection% reflectionSelection details
Rfree0.2702 3543 4.92 %RANDOM
Rwork0.2327 68469 --
all0.235 ---
obs-72012 98.14 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.2 Å / Solvent model: MASK BULK SOLVENT
Displacement parametersBiso mean: 111.481 Å2
Baniso -1Baniso -2Baniso -3
1--0.338 Å20 Å20 Å2
2---0.338 Å20 Å2
3---0.676 Å2
Refinement stepCycle: LAST / Resolution: 3.4→93.215 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms21818 0 68 0 21886
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.010.01222416
X-RAY DIFFRACTIONr_bond_other_d0.0010.01620468
X-RAY DIFFRACTIONr_angle_refined_deg2.161.81630374
X-RAY DIFFRACTIONr_angle_other_deg0.731.75747397
X-RAY DIFFRACTIONr_dihedral_angle_1_deg6.84652802
X-RAY DIFFRACTIONr_dihedral_angle_2_deg19.3845120
X-RAY DIFFRACTIONr_dihedral_angle_3_deg23.761103624
X-RAY DIFFRACTIONr_dihedral_angle_6_deg15.551101050
X-RAY DIFFRACTIONr_chiral_restr0.1020.23235
X-RAY DIFFRACTIONr_gen_planes_refined0.010.0226510
X-RAY DIFFRACTIONr_gen_planes_other0.0020.025006
X-RAY DIFFRACTIONr_nbd_refined0.2460.25902
X-RAY DIFFRACTIONr_symmetry_nbd_other0.2280.222538
X-RAY DIFFRACTIONr_nbtor_refined0.20.211361
X-RAY DIFFRACTIONr_symmetry_nbtor_other0.0910.212624
X-RAY DIFFRACTIONr_xyhbond_nbd_refined0.1540.2786
X-RAY DIFFRACTIONr_symmetry_xyhbond_nbd_other0.0430.27
X-RAY DIFFRACTIONr_metal_ion_refined0.2320.25
X-RAY DIFFRACTIONr_symmetry_nbd_refined0.260.2142
X-RAY DIFFRACTIONr_nbd_other0.2740.2418
X-RAY DIFFRACTIONr_symmetry_xyhbond_nbd_refined0.2840.224
X-RAY DIFFRACTIONr_xyhbond_nbd_other0.0510.22
X-RAY DIFFRACTIONr_mcbond_it4.7213.00711226
X-RAY DIFFRACTIONr_mcbond_other4.7213.00711226
X-RAY DIFFRACTIONr_mcangle_it7.9495.3914022
X-RAY DIFFRACTIONr_mcangle_other7.9485.38914023
X-RAY DIFFRACTIONr_scbond_it4.6523.34411190
X-RAY DIFFRACTIONr_scbond_other4.6523.34411191
X-RAY DIFFRACTIONr_scangle_it7.9496.0116352
X-RAY DIFFRACTIONr_scangle_other7.9496.0116353
X-RAY DIFFRACTIONr_lrange_it15.02336.95699077
X-RAY DIFFRACTIONr_lrange_other15.02236.95599077
X-RAY DIFFRACTIONr_ncsr_local_group_10.1590.0514717
X-RAY DIFFRACTIONr_ncsr_local_group_20.1740.0514328
X-RAY DIFFRACTIONr_ncsr_local_group_30.1570.0514620
X-RAY DIFFRACTIONr_ncsr_local_group_40.1680.0514465
X-RAY DIFFRACTIONr_ncsr_local_group_50.1790.0514179
X-RAY DIFFRACTIONr_ncsr_local_group_60.1720.0514349
X-RAY DIFFRACTIONr_ncsr_local_group_70.1740.0514453
X-RAY DIFFRACTIONr_ncsr_local_group_80.1780.0514451
X-RAY DIFFRACTIONr_ncsr_local_group_90.1840.0514230
X-RAY DIFFRACTIONr_ncsr_local_group_100.1750.0514220
X-RAY DIFFRACTIONr_ncsr_local_group_110.1710.0514529
X-RAY DIFFRACTIONr_ncsr_local_group_120.1690.0514404
X-RAY DIFFRACTIONr_ncsr_local_group_130.170.0514448
X-RAY DIFFRACTIONr_ncsr_local_group_140.1780.0514120
X-RAY DIFFRACTIONr_ncsr_local_group_150.1660.0514560
Refine LS restraints NCS
Ens-IDDom-IDAuth asym-IDRefine-IDTypeRms dev position (Å)Weight position
11AX-RAY DIFFRACTIONLocal ncs0.158880.05007
12BX-RAY DIFFRACTIONLocal ncs0.158880.05007
23AX-RAY DIFFRACTIONLocal ncs0.173520.05007
24CX-RAY DIFFRACTIONLocal ncs0.173520.05007
35AX-RAY DIFFRACTIONLocal ncs0.156560.05008
36DX-RAY DIFFRACTIONLocal ncs0.156560.05008
47AX-RAY DIFFRACTIONLocal ncs0.168030.05007
48EX-RAY DIFFRACTIONLocal ncs0.168030.05007
59AX-RAY DIFFRACTIONLocal ncs0.179040.05007
510FX-RAY DIFFRACTIONLocal ncs0.179040.05007
611BX-RAY DIFFRACTIONLocal ncs0.172330.05007
612CX-RAY DIFFRACTIONLocal ncs0.172330.05007
713BX-RAY DIFFRACTIONLocal ncs0.174370.05007
714DX-RAY DIFFRACTIONLocal ncs0.174370.05007
815BX-RAY DIFFRACTIONLocal ncs0.177650.05007
816EX-RAY DIFFRACTIONLocal ncs0.177650.05007
917BX-RAY DIFFRACTIONLocal ncs0.184150.05007
918FX-RAY DIFFRACTIONLocal ncs0.184150.05007
1019CX-RAY DIFFRACTIONLocal ncs0.174760.05007
1020DX-RAY DIFFRACTIONLocal ncs0.174760.05007
1121CX-RAY DIFFRACTIONLocal ncs0.170670.05007
1122EX-RAY DIFFRACTIONLocal ncs0.170670.05007
1223CX-RAY DIFFRACTIONLocal ncs0.169380.05007
1224FX-RAY DIFFRACTIONLocal ncs0.169380.05007
1325DX-RAY DIFFRACTIONLocal ncs0.169690.05007
1326EX-RAY DIFFRACTIONLocal ncs0.169690.05007
1427DX-RAY DIFFRACTIONLocal ncs0.177790.05007
1428FX-RAY DIFFRACTIONLocal ncs0.177790.05007
1529EX-RAY DIFFRACTIONLocal ncs0.166010.05007
1530FX-RAY DIFFRACTIONLocal ncs0.166010.05007
LS refinement shell

Refine-ID: X-RAY DIFFRACTION / Total num. of bins used: 20

Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRfactor allNum. reflection allFsc freeFsc work% reflection obs (%)WRfactor Rwork
3.4-3.4880.3432420.34350010.34353590.9130.91697.83540.317
3.488-3.5840.3212620.28748200.28952040.9320.94797.65560.25
3.584-3.6870.3022400.27947220.2850700.9340.94697.86980.239
3.687-3.8010.2742390.2546130.25149200.9520.9698.61790.208
3.801-3.9250.3182050.25945270.26147880.9360.95298.83040.222
3.925-4.0630.2792390.23343230.23546310.9510.96598.510.193
4.063-4.2160.2722350.2242260.22344960.9540.9799.22150.179
4.216-4.3880.2132040.19740450.19843010.9740.97698.7910.157
4.388-4.5820.2221830.19439140.19541520.9720.97798.67530.151
4.582-4.8050.2452300.19736530.239680.970.97797.85790.159
4.805-5.0640.2591510.21135560.21337790.9620.97498.09470.172
5.064-5.3710.2321930.19633210.19835740.970.97798.32120.162
5.371-5.740.2731610.23431760.23633910.9510.96698.40750.197
5.74-6.1980.3281670.23829450.24331660.9270.96598.29440.206
6.198-6.7870.3081430.23827310.24129210.9410.96398.3910.205
6.787-7.5830.2421450.19924600.20226620.9650.97597.85870.176
7.583-8.7460.221000.222010.20123720.970.97497.00670.185
8.746-10.6880.272890.20118820.20420210.9590.97697.5260.198
10.688-15.0170.251750.22414850.22516170.9620.97396.4750.225
15.017-93.2150.322400.3718680.3699790.9430.91292.74770.463
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)
13.18770.32050.2691.3843-0.20791.5916-0.05070.141-0.23430.06770.1187-0.57570.07810.4498-0.0680.0110.0314-0.05040.1505-0.08291.032195.672-53.024-33.1199
22.63750.9381-0.09392.97450.99421.5162-0.0177-0.67720.52730.5559-0.1044-0.148-0.15210.29820.12220.2458-0.0352-0.21490.2342-0.19611.213378.1923-18.0898-13.8458
32.28260.3124-0.25813.59910.04660.4786-0.109-0.15970.67620.05-0.03250.414-0.4805-0.12390.14150.56370.1746-0.38410.066-0.22931.64842.83650.9414-31.273
42.2221-0.05760.21692.3634-0.9411.58320.2491-0.9182-0.41760.9808-0.0807-0.25820.3252-0.1092-0.16840.8425-0.1819-0.31160.38730.18731.256268.88-78.0603-7.3731
50.70620.1025-0.3920.8938-0.48533.62030.0543-0.89970.20020.9204-0.22540.1206-0.0061-0.33320.1711.0214-0.2980.12251.3978-0.33251.223949.8798-43.09289.9816
62.25040.3898-0.8782.10060.24562.17910.0081-0.73150.48780.697-0.27570.9222-0.3201-0.7840.26770.46170.15560.16120.9823-0.71071.899414.867-25.8694-9.1554
Refinement TLS group
IDRefine-IDRefine TLS-IDSelectionAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1ALLA2 - 503
2X-RAY DIFFRACTION2ALLB2 - 502
3X-RAY DIFFRACTION3ALLC2 - 501
4X-RAY DIFFRACTION4ALLD2 - 501
5X-RAY DIFFRACTION5ALLE2 - 501
6X-RAY DIFFRACTION6ALLF2 - 501

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