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- PDB-2gm3: Crystal Structure of an Universal Stress Protein Family Protein f... -

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Basic information

Entry
Database: PDB / ID: 2gm3
TitleCrystal Structure of an Universal Stress Protein Family Protein from Arabidopsis Thaliana At3g01520 with AMP Bound
Componentsunknown protein
KeywordsSTRUCTURAL GENOMICS / UNKNOWN FUNCTION / At3g01520 / putative ethylene-responsive protein / USP Domain / nucleotide binding domain / AMP / PROTEIN STRUCTURE INITIATIVE / PSI / CENTER FOR EUKARYOTIC STRUCTURAL GENOMICS / CESG
Function / homology
Function and homology information


AMP binding / plasma membrane / cytosol
Similarity search - Function
Universal stress protein A family / UspA / Universal stress protein family / HUPs / Rossmann-like alpha/beta/alpha sandwich fold / Rossmann fold / 3-Layer(aba) Sandwich / Alpha Beta
Similarity search - Domain/homology
ADENOSINE MONOPHOSPHATE / Universal stress protein A-like protein
Similarity search - Component
Biological speciesArabidopsis thaliana (thale cress)
MethodX-RAY DIFFRACTION / SYNCHROTRON / SAD / Resolution: 2.461 Å
AuthorsBitto, E. / Wesenberg, G.E. / Phillips Jr., G.N. / Bingman, C.A. / Center for Eukaryotic Structural Genomics (CESG)
CitationJournal: Proteins / Year: 2015
Title: Crystal structure of the protein At3g01520, a eukaryotic universal stress protein-like protein from arabidopsis thaliana in complex with AMP.
Authors: Kim, D.O.J. / Bitto, E. / Bingman, C.A. / Kim, H.J. / Han, B.W. / Phillips, G.N.
History
DepositionApr 5, 2006Deposition site: RCSB / Processing site: RCSB
Revision 1.0Apr 18, 2006Provider: repository / Type: Initial release
Revision 1.1May 1, 2008Group: Version format compliance
Revision 1.2Jul 13, 2011Group: Advisory / Version format compliance
Revision 1.3Jun 8, 2016Group: Database references
Revision 1.4Oct 18, 2017Group: Refinement description / Category: software

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

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Assembly

Deposited unit
A: unknown protein
B: unknown protein
C: unknown protein
D: unknown protein
E: unknown protein
F: unknown protein
hetero molecules


Theoretical massNumber of molelcules
Total (without water)120,23512
Polymers118,1526
Non-polymers2,0836
Water23413
1
A: unknown protein
B: unknown protein
hetero molecules


Theoretical massNumber of molelcules
Total (without water)40,0784
Polymers39,3842
Non-polymers6942
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area3360 Å2
ΔGint-11 kcal/mol
Surface area15340 Å2
MethodPISA
2
C: unknown protein
D: unknown protein
hetero molecules


Theoretical massNumber of molelcules
Total (without water)40,0784
Polymers39,3842
Non-polymers6942
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area3350 Å2
ΔGint-10 kcal/mol
Surface area15390 Å2
MethodPISA
3
E: unknown protein
F: unknown protein
hetero molecules


Theoretical massNumber of molelcules
Total (without water)40,0784
Polymers39,3842
Non-polymers6942
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
4
C: unknown protein
E: unknown protein
hetero molecules


Theoretical massNumber of molelcules
Total (without water)40,0784
Polymers39,3842
Non-polymers6942
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area3020 Å2
ΔGint-20 kcal/mol
Surface area15740 Å2
MethodPISA
5
A: unknown protein
hetero molecules

D: unknown protein
hetero molecules


Theoretical massNumber of molelcules
Total (without water)40,0784
Polymers39,3842
Non-polymers6942
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
crystal symmetry operation1_665x+1,y+1,z1
Buried area3030 Å2
ΔGint-18 kcal/mol
Surface area15680 Å2
MethodPISA
6
B: unknown protein
hetero molecules

F: unknown protein
hetero molecules


Theoretical massNumber of molelcules
Total (without water)40,0784
Polymers39,3842
Non-polymers6942
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
crystal symmetry operation1_446x-1,y-1,z+11
Buried area3040 Å2
ΔGint-21 kcal/mol
Surface area15870 Å2
MethodPISA
Unit cell
Length a, b, c (Å)63.351, 65.662, 73.014
Angle α, β, γ (deg.)75.45, 75.04, 66.11
Int Tables number1
Space group name H-MP1
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11A
21B
31C
41D
51E
61F
71A
81B
91C
101D
111E
121F
12A
22B
32C
42D
52D
62F

NCS domain segments:

Refine code: 2

Dom-IDComponent-IDEns-IDBeg label comp-IDEnd label comp-IDAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
111PROVALAA5 - 555 - 55
211PROVALBB5 - 555 - 55
311PROVALCC5 - 555 - 55
411PROVALDD5 - 555 - 55
511PROVALEE5 - 555 - 55
611PROVALFF5 - 555 - 55
721GLYASPAA85 - 17485 - 174
821GLYASPBB85 - 17485 - 174
921GLYASPCC85 - 17485 - 174
1021GLYASPDD85 - 17485 - 174
1121GLYASPEE85 - 17485 - 174
1221GLYASPFF85 - 17485 - 174
112SERMSEAA66 - 7766 - 77
212SERMSEBB66 - 7766 - 77
312SERMSECC66 - 7766 - 77
412SERMSEDD66 - 7766 - 77
512SERMSEDD66 - 7766 - 77
612SERMSEFF66 - 7766 - 77

NCS ensembles :
ID
1
2
DetailsThe biological unit is a dimer. There are 3 biological units in the asymmetric unit (chains A & B and chains C & D and chains E & F).

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Components

#1: Protein
unknown protein


Mass: 19691.934 Da / Num. of mol.: 6
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Arabidopsis thaliana (thale cress) / Gene: At3g01520 / Plasmid: PVP 16 / Production host: Escherichia coli (E. coli) / Strain (production host): BL834 P(RARE2) / References: UniProt: Q8LGG8
#2: Chemical
ChemComp-AMP / ADENOSINE MONOPHOSPHATE / Adenosine monophosphate


Mass: 347.221 Da / Num. of mol.: 6 / Source method: obtained synthetically / Formula: C10H14N5O7P / Comment: AMP*YM
#3: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 13 / 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: 2.24 Å3/Da / Density % sol: 45 %
Crystal growTemperature: 293 K / Method: vapor diffusion, hanging drop
Details: PROTEIN SOLUTION (10 MG/ML PROTEIN, 0.050 M SODIUM CHLORIDE, 0.003 M SODIUM AZIDE, 0.0003 M TCEP, 0.005 MES PH 7.0) MIXED IN A 1:1 RATIO WITH THE WELL SOLUTION (18% PEG 2K, 5% DMSO, 0.10 M ...Details: PROTEIN SOLUTION (10 MG/ML PROTEIN, 0.050 M SODIUM CHLORIDE, 0.003 M SODIUM AZIDE, 0.0003 M TCEP, 0.005 MES PH 7.0) MIXED IN A 1:1 RATIO WITH THE WELL SOLUTION (18% PEG 2K, 5% DMSO, 0.10 M PIPES PH 6.5) Crystals cryo-protected with 20% PEG 2K, 0.1 M TRIS PH 8, and a final concentration of 20 % Ethylene glycol, temperature 293K, VAPOR DIFFUSION, HANGING DROP

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Data collection

Diffraction
IDMean temperature (K)Crystal-ID
11001
21
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 22-ID / Wavelength: 0.97934 Å
DetectorType: MARMOSAIC 300 mm CCD / Detector: CCD / Date: Jun 13, 2005
Details: HORIZONTAL SAGITALLY FOCUSING 2ND BENT MONOCHROMATOR CRYSTAL, VERTICAL BENT FOCUSING MIRROR
RadiationMonochromator: CRYOGENICALLY COOLED SI (220) DOUBLE BOUNCE / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.97934 Å / Relative weight: 1
ReflectionResolution: 2.5→40.7 Å / Num. obs: 36006 / % possible obs: 97.2 % / Redundancy: 3.7 % / Rmerge(I) obs: 0.058 / Χ2: 1.305 / Net I/σ(I): 9.913
Reflection shellResolution: 2.5→2.59 Å / Redundancy: 3 % / Rmerge(I) obs: 0.239 / Mean I/σ(I) obs: 4.406 / Num. unique all: 3309 / Χ2: 1.131 / % possible all: 89.4

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Phasing

Phasing dmFOM : 0.66 / FOM acentric: 0.66 / FOM centric: 0 / Reflection: 36473 / Reflection acentric: 36473 / Reflection centric: 0
Phasing dm shell
Resolution (Å)FOM FOM acentricReflectionReflection acentric
7-69.5590.940.9415921592
4.4-70.890.8949304930
3.5-4.40.850.8562186218
3.1-3.50.690.6962856285
2.6-3.10.520.521115811158
2.5-2.60.440.4462906290

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Processing

Software
NameVersionClassificationNB
DENZOdata reduction
SCALEPACKdata scaling
RESOLVE2.06phasing
REFMAC5.2.0005refinement
PDB_EXTRACT1.701data extraction
SHELXEmodel building
RefinementMethod to determine structure: SAD / Resolution: 2.461→40.7 Å / Cor.coef. Fo:Fc: 0.938 / Cor.coef. Fo:Fc free: 0.914 / WRfactor Rfree: 0.288 / WRfactor Rwork: 0.24 / SU B: 18.763 / SU ML: 0.229 / TLS residual ADP flag: LIKELY RESIDUAL / Cross valid method: THROUGHOUT / ESU R: 0.651 / ESU R Free: 0.304 / Stereochemistry target values: MAXIMUM LIKELIHOOD / Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS
RfactorNum. reflection% reflectionSelection details
Rfree0.2607 1800 4.999 %Random
Rwork0.2185 ---
all0.221 ---
obs0.22065 36006 96.822 %-
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: 60.244 Å2
Baniso -1Baniso -2Baniso -3
1--0.171 Å2-0.933 Å21.28 Å2
2--0.245 Å2-0.48 Å2
3---0.262 Å2
Refinement stepCycle: LAST / Resolution: 2.461→40.7 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms7084 0 138 13 7235
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0160.0227380
X-RAY DIFFRACTIONr_angle_refined_deg1.5011.9779995
X-RAY DIFFRACTIONr_dihedral_angle_1_deg5.6755889
X-RAY DIFFRACTIONr_dihedral_angle_2_deg34.46324.007307
X-RAY DIFFRACTIONr_dihedral_angle_3_deg17.647151286
X-RAY DIFFRACTIONr_dihedral_angle_4_deg21.7111547
X-RAY DIFFRACTIONr_chiral_restr0.3740.21146
X-RAY DIFFRACTIONr_gen_planes_refined0.0050.025395
X-RAY DIFFRACTIONr_nbd_refined0.2220.23463
X-RAY DIFFRACTIONr_nbtor_refined0.310.25031
X-RAY DIFFRACTIONr_xyhbond_nbd_refined0.160.2257
X-RAY DIFFRACTIONr_symmetry_vdw_refined0.1960.2111
X-RAY DIFFRACTIONr_symmetry_hbond_refined0.2620.212
X-RAY DIFFRACTIONr_mcbond_it0.5731.54623
X-RAY DIFFRACTIONr_mcangle_it0.94227371
X-RAY DIFFRACTIONr_scbond_it1.44533097
X-RAY DIFFRACTIONr_scangle_it2.184.52622
Refine LS restraints NCS

Refine-ID: X-RAY DIFFRACTION

Ens-IDDom-IDAuth asym-IDNumberTypeRms dev position (Å)Weight position
11A532TIGHT POSITIONAL0.070.05
12B532TIGHT POSITIONAL0.060.05
13C532TIGHT POSITIONAL0.050.05
14D532TIGHT POSITIONAL0.060.05
15E532TIGHT POSITIONAL0.040.05
16F532TIGHT POSITIONAL0.050.05
11A500MEDIUM POSITIONAL0.480.5
12B500MEDIUM POSITIONAL0.610.5
13C500MEDIUM POSITIONAL0.380.5
14D500MEDIUM POSITIONAL0.620.5
15E500MEDIUM POSITIONAL0.420.5
16F500MEDIUM POSITIONAL0.720.5
11A532TIGHT THERMAL0.120.5
12B532TIGHT THERMAL0.10.5
13C532TIGHT THERMAL0.110.5
14D532TIGHT THERMAL0.10.5
15E532TIGHT THERMAL0.10.5
16F532TIGHT THERMAL0.090.5
11A500MEDIUM THERMAL0.82
12B500MEDIUM THERMAL0.662
13C500MEDIUM THERMAL0.732
14D500MEDIUM THERMAL0.782
15E500MEDIUM THERMAL0.572
16F500MEDIUM THERMAL0.642
21A47TIGHT POSITIONAL0.050.05
22B47TIGHT POSITIONAL0.050.05
23C47TIGHT POSITIONAL0.040.05
24D47TIGHT POSITIONAL0.030.05
22E47TIGHT POSITIONAL0.030.05
26F47TIGHT POSITIONAL0.070.05
21A47MEDIUM POSITIONAL0.590.5
22B47MEDIUM POSITIONAL0.740.5
23C47MEDIUM POSITIONAL0.710.5
24D47MEDIUM POSITIONAL0.740.5
22E47MEDIUM POSITIONAL0.740.5
26F47MEDIUM POSITIONAL0.930.5
21A47TIGHT THERMAL0.240.5
22B47TIGHT THERMAL0.20.5
23C47TIGHT THERMAL0.080.5
24D47TIGHT THERMAL0.110.5
22E47TIGHT THERMAL0.110.5
26F47TIGHT THERMAL0.220.5
21A47MEDIUM THERMAL0.742
22B47MEDIUM THERMAL1.252
23C47MEDIUM THERMAL0.452
24D47MEDIUM THERMAL0.732
22E47MEDIUM THERMAL0.732
26F47MEDIUM THERMAL12
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 all% reflection obs (%)
2.461-2.5250.3281220.29322340.295282783.339
2.525-2.5940.3511310.27623300.28263293.503
2.594-2.6690.3651440.2623530.266257796.896
2.669-2.7510.2831110.25723980.258255498.238
2.751-2.8420.2871120.2523040.252246298.132
2.842-2.9410.2731060.24222310.243237198.566
2.941-3.0520.31050.22621480.229229098.384
3.052-3.1760.2371040.2320810.23221898.512
3.176-3.3170.2891190.21519490.219208699.137
3.317-3.4790.277880.22619180.228203298.72
3.479-3.6660.236940.22318000.224191199.11
3.666-3.8880.261870.21216970.214181298.455
3.888-4.1560.228840.216070.201171098.889
4.156-4.4870.222790.17514760.177157698.668
4.487-4.9130.199730.1813630.181146398.154
4.913-5.490.282660.21612480.219132499.245
5.49-6.3330.312530.24211030.244116199.569
6.333-7.740.29570.2299210.23398898.988
7.74-10.8790.193430.1747020.17576996.879
10.879-69.5050.352220.2883430.29142585.882
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)
15.41081.4193-0.15234.4523-0.24754.11960.04530.2363-0.04130.144-0.2015-0.1017-0.23050.0560.1563-0.2876-0.025-0.0661-0.3186-0.0079-0.28826.28818.6961.014
25.04510.1191-0.84686.966-0.40974.85720.0519-0.44-0.66230.7369-0.13450.45620.4284-0.51620.0827-0.1569-0.19890.1031-0.0546-0.05-0.112110.5434.43472.917
36.77470.5106-0.01323.9315-0.28414.61360.3270.18910.1657-0.0663-0.282-0.6015-0.3030.3792-0.0449-0.26260.03420.0601-0.30140.0638-0.2095-5.702-13.02739.772
46.79941.1305-1.37014.19350.01244.93590.3092-0.3275-0.15210.1691-0.22820.3321-0.0468-0.462-0.0811-0.30980.0132-0.0048-0.22280.0176-0.2983-28.589-18.5747.871
56.49431.6583-0.16157.5561-0.07164.9389-0.09340.1138-0.22220.66770.54430.28760.4132-0.3493-0.451-0.08450.0662-0.0081-0.14790.0604-0.269127.9769.5723.521
63.2234-0.1391-0.373210.2477-0.86834.85170.09760.00970.61780.25020.5819-1.917-0.51550.517-0.6795-0.1747-0.0150.0693-0.0512-0.29950.32545.48227.34620.954
Refinement TLS group

Refine-ID: X-RAY DIFFRACTION / Selection: ALL

IDRefine TLS-IDAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
11AA5 - 555 - 55
21AA80 - 17480 - 174
32BB5 - 555 - 55
42BB85 - 17485 - 174
53CC5 - 555 - 55
63CC80 - 17480 - 174
74DD5 - 555 - 55
84DD80 - 17480 - 174
95EE5 - 555 - 55
105EE80 - 17480 - 174
116FF5 - 555 - 55
126FF79 - 17479 - 174

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