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- PDB-5kod: Crystal Structure of GH3.5 Acyl Acid Amido Synthetase from Arabid... -

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Entry
Database: PDB / ID: 5kod
TitleCrystal Structure of GH3.5 Acyl Acid Amido Synthetase from Arabidopsis thaliana
ComponentsIndole-3-acetic acid-amido synthetase GH3.5
KeywordsLIGASE / ANL / adenylating enzyme / acyl acid amido synthetase / adenylation ligase
Function / homology
Function and homology information


indole-3-acetic acid amido synthetase activity / positive regulation of camalexin biosynthetic process / camalexin biosynthetic process / Ligases; Forming carbon-nitrogen bonds; Acid-amino-acid ligases (peptide synthases) / response to auxin
Similarity search - Function
GH3 family / GH3 auxin-responsive promoter
Similarity search - Domain/homology
ADENOSINE MONOPHOSPHATE / 1H-INDOL-3-YLACETIC ACID / Indole-3-acetic acid-amido synthetase GH3.5
Similarity search - Component
Biological speciesArabidopsis thaliana (thale cress)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.202 Å
AuthorsJez, J.M. / Westfall, C.S. / Zubieta, C.
Funding support United States, 1items
OrganizationGrant numberCountry
National Science Foundation (NSF, United States)MCB-1157771 United States
CitationJournal: Proc. Natl. Acad. Sci. U.S.A. / Year: 2016
Title: Arabidopsis thaliana GH3.5 acyl acid amido synthetase mediates metabolic crosstalk in auxin and salicylic acid homeostasis.
Authors: Westfall, C.S. / Sherp, A.M. / Zubieta, C. / Alvarez, S. / Schraft, E. / Marcellin, R. / Ramirez, L. / Jez, J.M.
History
DepositionJun 30, 2016Deposition site: RCSB / Processing site: RCSB
Revision 1.0Nov 16, 2016Provider: repository / Type: Initial release
Revision 1.1Nov 30, 2016Group: Database references
Revision 1.2Dec 14, 2016Group: Database references
Revision 1.3Sep 27, 2017Group: Author supporting evidence / Category: pdbx_audit_support / Item: _pdbx_audit_support.funding_organization
Revision 1.4Nov 27, 2019Group: Author supporting evidence / Category: pdbx_audit_support / Item: _pdbx_audit_support.funding_organization
Revision 1.5Oct 4, 2023Group: 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
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Indole-3-acetic acid-amido synthetase GH3.5
B: Indole-3-acetic acid-amido synthetase GH3.5
C: Indole-3-acetic acid-amido synthetase GH3.5
D: Indole-3-acetic acid-amido synthetase GH3.5
hetero molecules


Theoretical massNumber of molelcules
Total (without water)279,71414
Polymers277,4324
Non-polymers2,28210
Water18,0331001
1
A: Indole-3-acetic acid-amido synthetase GH3.5
hetero molecules


Theoretical massNumber of molelcules
Total (without water)69,8803
Polymers69,3581
Non-polymers5222
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA
2
B: Indole-3-acetic acid-amido synthetase GH3.5
hetero molecules


Theoretical massNumber of molelcules
Total (without water)70,0735
Polymers69,3581
Non-polymers7154
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA
3
C: Indole-3-acetic acid-amido synthetase GH3.5
hetero molecules


Theoretical massNumber of molelcules
Total (without water)69,8803
Polymers69,3581
Non-polymers5222
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA
4
D: Indole-3-acetic acid-amido synthetase GH3.5
hetero molecules


Theoretical massNumber of molelcules
Total (without water)69,8803
Polymers69,3581
Non-polymers5222
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA
Unit cell
Length a, b, c (Å)91.609, 143.523, 102.322
Angle α, β, γ (deg.)90.00, 114.72, 90.00
Int Tables number4
Space group name H-MP1211

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Components

#1: Protein
Indole-3-acetic acid-amido synthetase GH3.5 / Auxin-responsive GH3-like protein 5 / AtGH3-5


Mass: 69358.086 Da / Num. of mol.: 4
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Arabidopsis thaliana (thale cress) / Gene: GH3.5, At4g27260, M4I22.70 / Production host: Escherichia coli (E. coli) / Strain (production host): Rosetta (DE3)
References: UniProt: O81829, Ligases; Forming carbon-nitrogen bonds; Acid-amino-acid ligases (peptide synthases)
#2: Chemical
ChemComp-AMP / ADENOSINE MONOPHOSPHATE


Mass: 347.221 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: C10H14N5O7P / Comment: AMP*YM
#3: Chemical
ChemComp-IAC / 1H-INDOL-3-YLACETIC ACID / INDOLE ACETIC ACID


Mass: 175.184 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: C10H9NO2 / Comment: hormone*YM
#4: Chemical ChemComp-SO4 / SULFATE ION


Mass: 96.063 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: SO4
#5: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 1001 / 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: 2.2 Å3/Da / Density % sol: 44.15 %
Crystal growTemperature: 277 K / Method: vapor diffusion, hanging drop / pH: 9.5
Details: 20% PEG-8000 0.1 M N-cyclohexyl-2-aminoethanesulfonic acid, pH 9.5 2 mM MgCl2 5 mM tris(2-carboxyethyl)phosphine 5 mM IAA 5 mM AMP

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: ESRF / Beamline: ID23-2 / Wavelength: 0.98 Å
DetectorType: MARMOSAIC 225 mm CCD / Detector: CCD / Date: Jun 11, 2012
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.98 Å / Relative weight: 1
ReflectionResolution: 2.2→42.2 Å / Num. obs: 108566 / % possible obs: 89.5 % / Observed criterion σ(F): 0 / Redundancy: 2.26 % / Rsym value: 0.093 / Net I/σ(I): 11.2

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Processing

Software
NameVersionClassification
PHENIX1.9_1692refinement
XDSdata reduction
XSCALEdata scaling
PHASERphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 4EQL
Resolution: 2.202→42.164 Å / SU ML: 0.28 / Cross valid method: FREE R-VALUE / σ(F): 1.99 / Phase error: 25 / Stereochemistry target values: MLHL
RfactorNum. reflection% reflection
Rfree0.2447 5419 4.99 %
Rwork0.2048 --
obs0.2068 108566 89.53 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL
Refinement stepCycle: LAST / Resolution: 2.202→42.164 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms18129 0 154 1001 19284
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.00818787
X-RAY DIFFRACTIONf_angle_d1.16725519
X-RAY DIFFRACTIONf_dihedral_angle_d16.2147073
X-RAY DIFFRACTIONf_chiral_restr0.0442876
X-RAY DIFFRACTIONf_plane_restr0.0053240
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
2.2019-2.22690.31051090.27222073X-RAY DIFFRACTION54
2.2269-2.25310.33421430.26072759X-RAY DIFFRACTION73
2.2531-2.28050.31131750.2553258X-RAY DIFFRACTION84
2.2805-2.30940.30671850.26843594X-RAY DIFFRACTION95
2.3094-2.33980.36821770.2973350X-RAY DIFFRACTION87
2.3398-2.37180.37171810.28553371X-RAY DIFFRACTION88
2.3718-2.40570.26131970.23513744X-RAY DIFFRACTION98
2.4057-2.44160.28531950.22263770X-RAY DIFFRACTION98
2.4416-2.47980.26621960.23123734X-RAY DIFFRACTION98
2.4798-2.52040.30532030.22333749X-RAY DIFFRACTION98
2.5204-2.56390.28342000.21633772X-RAY DIFFRACTION98
2.5639-2.61050.23881990.22423783X-RAY DIFFRACTION99
2.6105-2.66070.2781900.22841748X-RAY DIFFRACTION93
2.6607-2.7150.4051320.2892614X-RAY DIFFRACTION73
2.715-2.7740.33151630.27273163X-RAY DIFFRACTION82
2.774-2.83850.28961930.21863720X-RAY DIFFRACTION97
2.8385-2.90950.26772000.21243760X-RAY DIFFRACTION99
2.9095-2.98820.26871980.20883802X-RAY DIFFRACTION99
2.9882-3.07610.25571980.20593772X-RAY DIFFRACTION99
3.0761-3.17530.25471950.19973797X-RAY DIFFRACTION98
3.1753-3.28880.22881980.19733766X-RAY DIFFRACTION98
3.2888-3.42040.21692020.18263793X-RAY DIFFRACTION98
3.4204-3.5760.24161990.18753740X-RAY DIFFRACTION98
3.576-3.76440.19731960.17213760X-RAY DIFFRACTION98
3.7644-4.00010.22451960.16983739X-RAY DIFFRACTION97
4.0001-4.30870.19591930.1583766X-RAY DIFFRACTION97
4.3087-4.74170.18561970.15713744X-RAY DIFFRACTION98
4.7417-5.42660.18532010.17073807X-RAY DIFFRACTION98
5.4266-6.83230.22742020.20993837X-RAY DIFFRACTION99
6.8323-42.17180.23232060.22263862X-RAY DIFFRACTION98
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)
12.68830.54660.2531.6233-0.4610.7868-0.1488-0.24030.4140.2286-0.0306-0.1541-0.26040.09360.08990.4033-0.0228-0.0450.2527-0.05010.201336.558111.092366.4659
20.85890.34490.1951.0526-0.03920.91740.0202-0.0308-0.01420.08640.00960.02760.0472-0.0436-0.03560.25420.0429-0.0030.18240.00460.149620.706492.500759.1341
31.683-0.27930.08741.77240.44392.8190.0760.1726-0.0551-0.3078-0.0561-0.16450.10630.1429-0.01770.33730.04880.03870.19160.01570.232137.599679.750638.7488
44.90221.07240.25952.17250.49251.0813-0.06130.5433-0.35650.06640.1027-0.09010.38910.2415-0.08550.31530.05150.07440.2084-0.06950.260813.416884.266887.2626
52.9541-2.2844-0.42511.98220.97671.8049-0.0837-0.34430.3433-0.02530.2353-0.4398-0.16120.3128-0.16120.1922-0.01710.00580.2471-0.0220.2816.1077101.526693.9808
61.884-0.4112-0.29992.6422-0.12721.1434-0.03380.3357-0.1524-0.2173-0.08760.16160.0957-0.1430.1170.2298-0.03440.03520.2865-0.07450.1626-6.740191.768882.6995
72.7805-0.3142-0.69420.84560.23841.06570.05650.55950.1096-0.1414-0.08670.0654-0.0438-0.28770.03510.2395-0.002-0.00250.31770.00340.178-16.1525103.289584.3771
83.6536-1.0889-0.26221.91710.25972.1974-0.204-0.481-0.22960.40630.0497-0.08120.13360.00970.06040.28530.03230.04990.20930.00080.21923.536686.4844101.3604
92.1747-0.5973-0.66871.86290.63140.95-0.2599-0.2928-0.10680.20280.1525-0.07850.17280.10470.12250.26460.0450.01420.18450.01590.16774.235690.0539101.953
102.6131-0.08750.53812.9031.07732.6677-0.0215-0.18440.27320.62550.0724-0.18350.17180.088-0.04170.27880.0273-0.05030.189-0.01480.1847-9.0378118.1195116.3638
111.6529-0.5978-1.36162.23411.52882.6688-0.02090.06720.0736-0.02660.0738-0.1616-0.142-0.0076-0.0820.20630.0274-0.03640.1370.00560.1416-9.0847118.0017106.6177
121.73160.518-0.72912.8918-1.51563.8732-0.0633-0.1365-0.22160.53990.1063-0.20740.33320.0826-0.02810.39170.0239-0.0820.2023-0.03620.1872-13.6452108.8295118.3718
131.1689-4.5208-4.44432.00011.99982.0002-0.7208-2.17321.99073.4351-0.2613-1.742-1.67320.8830.96710.86820.0921-0.29240.47870.06160.32313.0417119.197292.0036
142.0710.53730.54960.8130.27140.91870.04830.1175-0.0980.00910.0504-0.06860.1030.1051-0.11670.26850.0262-0.00160.1574-0.050.2041-45.495863.021483.1774
152.0641-0.47810.02520.27310.49470.98090.09650.0882-0.668-0.04140.0659-0.5890.25720.2048-0.18020.47450.0526-0.0910.2829-0.10620.487-31.483853.827990.3216
163.9405-1.14960.19621.1064-0.79331.64250.2881-0.4744-0.5124-0.1336-0.11370.04540.2480.3788-0.14350.33950.0523-0.09680.4114-0.07340.4104-15.529158.8926103.5606
171.5889-0.15040.73510.1257-0.0381.37880.0667-0.11940.2312-0.0798-0.0211-0.0183-0.10030.0836-0.09480.2675-0.0095-0.0180.1691-0.05830.2767-42.056675.131491.2725
181.46750.13560.28772.26810.07294.01310.0204-0.0109-0.13790.42080.1819-0.20990.2288-0.0556-0.18350.36250.0349-0.0770.1702-0.02350.3035-46.114465.7723119.3494
192.3651-0.1332-1.46234.16661.50832.20490.40650.18410.18660.1858-0.22340.1506-0.115-0.0264-0.28840.4280.0757-0.1170.3315-0.05790.3477-44.611878.5439121.7385
201.63120.25710.80140.794-0.40511.43730.008-0.3082-0.15920.18440.12240.2640.0528-0.4476-0.13250.27910.04510.07470.39090.06010.3035-24.6765107.118644.2625
211.21590.40840.01661.0761-0.45690.71070.0069-0.10060.03760.06740.04970.0954-0.0546-0.0652-0.03820.23920.04690.00280.2240.0020.16-4.876117.445437.2926
221.2803-0.0390.27641.77-1.09732.8628-0.1221-0.2221-0.39480.07190.17970.15770.1456-0.2549-0.1190.25390.0170.01230.25190.11070.262-18.530794.737941.1572
231.5147-0.2124-0.53612.03151.18372.7179-0.0340.1748-0.2646-0.05330.03650.01930.2584-0.0433-0.00910.24670.0003-0.04160.1883-0.01790.2298-8.8182103.163911.1269
242.0903-0.4655-0.10663.98630.72892.82740.09050.0684-0.12820.1738-0.07570.07750.0896-0.15660.03520.2098-0.0389-0.06210.1708-0.02020.2357-6.8454106.364213.2821
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection details
1X-RAY DIFFRACTION1chain 'A' and (resid 18 through 91 )
2X-RAY DIFFRACTION2chain 'A' and (resid 92 through 446 )
3X-RAY DIFFRACTION3chain 'A' and (resid 447 through 608 )
4X-RAY DIFFRACTION4chain 'B' and (resid 16 through 66 )
5X-RAY DIFFRACTION5chain 'B' and (resid 67 through 123 )
6X-RAY DIFFRACTION6chain 'B' and (resid 124 through 160 )
7X-RAY DIFFRACTION7chain 'B' and (resid 161 through 347 )
8X-RAY DIFFRACTION8chain 'B' and (resid 348 through 396 )
9X-RAY DIFFRACTION9chain 'B' and (resid 397 through 459 )
10X-RAY DIFFRACTION10chain 'B' and (resid 460 through 512 )
11X-RAY DIFFRACTION11chain 'B' and (resid 513 through 552 )
12X-RAY DIFFRACTION12chain 'B' and (resid 553 through 602 )
13X-RAY DIFFRACTION13chain 'B' and (resid 603 through 604 )
14X-RAY DIFFRACTION14chain 'C' and (resid 16 through 161 )
15X-RAY DIFFRACTION15chain 'C' and (resid 162 through 206 )
16X-RAY DIFFRACTION16chain 'C' and (resid 207 through 302 )
17X-RAY DIFFRACTION17chain 'C' and (resid 303 through 446 )
18X-RAY DIFFRACTION18chain 'C' and (resid 447 through 552 )
19X-RAY DIFFRACTION19chain 'C' and (resid 553 through 601 )
20X-RAY DIFFRACTION20chain 'D' and (resid 16 through 161 )
21X-RAY DIFFRACTION21chain 'D' and (resid 162 through 401 )
22X-RAY DIFFRACTION22chain 'D' and (resid 402 through 446 )
23X-RAY DIFFRACTION23chain 'D' and (resid 447 through 573 )
24X-RAY DIFFRACTION24chain 'D' and (resid 574 through 606 )

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