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- PDB-6o63: Crystal Structure of Arabidopsis thaliana Spermidine Synthase iso... -

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Basic information

Entry
Database: PDB / ID: 6o63
TitleCrystal Structure of Arabidopsis thaliana Spermidine Synthase isoform 1 (AtSPDS1)
ComponentsSpermidine synthase 1
KeywordsTRANSFERASE / Polyamine metabolism / Putrescine Biosynthesis / dc-SAM / MTA
Function / homology
Function and homology information


spermidine synthase / spermidine synthase activity / spermidine biosynthetic process / cytosol
Similarity search - Function
Spermidine/spermine synthase, eukaryotes / Spermidine synthase, tetramerisation domain / Polyamine biosynthesis domain, conserved site / Polyamine biosynthesis (PABS) domain signature. / Spermidine/spermine synthases / Polyamine biosynthesis domain / Spermidine synthase, tetramerisation domain / Spermidine synthase, tetramerisation domain superfamily / Spermidine synthase tetramerisation domain / Polyamine biosynthesis (PABS) domain profile. ...Spermidine/spermine synthase, eukaryotes / Spermidine synthase, tetramerisation domain / Polyamine biosynthesis domain, conserved site / Polyamine biosynthesis (PABS) domain signature. / Spermidine/spermine synthases / Polyamine biosynthesis domain / Spermidine synthase, tetramerisation domain / Spermidine synthase, tetramerisation domain superfamily / Spermidine synthase tetramerisation domain / Polyamine biosynthesis (PABS) domain profile. / Spermine/spermidine synthase domain / Spermidine Synthase; Chain: A, domain 2 / Vaccinia Virus protein VP39 / Roll / S-adenosyl-L-methionine-dependent methyltransferase superfamily / Rossmann fold / 3-Layer(aba) Sandwich / Mainly Beta / Alpha Beta
Similarity search - Domain/homology
DI(HYDROXYETHYL)ETHER / Spermidine synthase 1
Similarity search - Component
Biological speciesArabidopsis thaliana (thale cress)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 1.8 Å
AuthorsSekula, B. / Dauter, Z.
Funding support United States, 1items
OrganizationGrant numberCountry
National Institutes of Health/National Cancer Institute (NIH/NCI)The Intramural Research Program United States
CitationJournal: Front Plant Sci / Year: 2019
Title: Spermidine Synthase (SPDS) Undergoes Concerted Structural Rearrangements Upon Ligand Binding - A Case Study of the Two SPDS Isoforms FromArabidopsis thaliana.
Authors: Sekula, B. / Dauter, Z.
History
DepositionMar 5, 2019Deposition site: RCSB / Processing site: RCSB
Revision 1.0Jun 5, 2019Provider: repository / Type: Initial release
Revision 1.1Dec 4, 2019Group: Author supporting evidence / Category: pdbx_audit_support / Item: _pdbx_audit_support.funding_organization
Revision 1.2Mar 13, 2024Group: Data collection / Database references / Category: chem_comp_atom / chem_comp_bond / database_2
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

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Assembly

Deposited unit
A: Spermidine synthase 1
B: Spermidine synthase 1
C: Spermidine synthase 1
D: Spermidine synthase 1
hetero molecules


Theoretical massNumber of molelcules
Total (without water)148,43614
Polymers147,4514
Non-polymers98510
Water17,763986
1
A: Spermidine synthase 1
B: Spermidine synthase 1
hetero molecules


Theoretical massNumber of molelcules
Total (without water)74,2187
Polymers73,7252
Non-polymers4925
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area5650 Å2
ΔGint-35 kcal/mol
Surface area21280 Å2
MethodPISA
2
C: Spermidine synthase 1
D: Spermidine synthase 1
hetero molecules


Theoretical massNumber of molelcules
Total (without water)74,2187
Polymers73,7252
Non-polymers4925
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area4870 Å2
ΔGint-31 kcal/mol
Surface area20740 Å2
MethodPISA
Unit cell
Length a, b, c (Å)89.536, 76.338, 90.192
Angle α, β, γ (deg.)90.00, 104.68, 90.00
Int Tables number4
Space group name H-MP1211

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Components

#1: Protein
Spermidine synthase 1 / / SPDSY 1 / Putrescine aminopropyltransferase 1


Mass: 36862.645 Da / Num. of mol.: 4
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Arabidopsis thaliana (thale cress) / Tissue: Leaves / Gene: SPDSYN1, At1g23820, F5O8.38 / Production host: Escherichia coli BL21(DE3) (bacteria) / Strain (production host): BL21(DE3) / Tissue (production host): Leaves / References: UniProt: Q9ZUB3, spermidine synthase
#2: Chemical
ChemComp-PEG / DI(HYDROXYETHYL)ETHER / Diethylene glycol


Mass: 106.120 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: C4H10O3
#3: Chemical
ChemComp-GOL / GLYCEROL / GLYCERIN / PROPANE-1,2,3-TRIOL / Glycerol


Mass: 92.094 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: C3H8O3
#4: Chemical ChemComp-SO4 / SULFATE ION / Sulfate


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


Mass: 18.015 Da / Num. of mol.: 986 / 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: 2.02 Å3/Da / Density % sol: 39.17 %
Crystal growTemperature: 292 K / Method: vapor diffusion, sitting drop / pH: 5.5
Details: 0.2 M ammonium sulfate, 0.1 M BIS-TRIS, 25% PEG 3350, cryoprotection 25% glycerol

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Data collection

DiffractionMean temperature: 100 K / Serial crystal experiment: N
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 22-BM / Wavelength: 1 Å
DetectorType: RAYONIX MX300-HS / Detector: CCD / Date: Nov 1, 2018
RadiationMonochromator: Si (111) / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 1 Å / Relative weight: 1
ReflectionResolution: 1.8→44.6 Å / Num. obs: 95958 / % possible obs: 95.4 % / Observed criterion σ(I): -3 / Redundancy: 4.1 % / CC1/2: 0.999 / Rmerge(I) obs: 0.055 / Net I/σ(I): 14.1
Reflection shellResolution: 1.8→1.86 Å / Redundancy: 4 % / Rmerge(I) obs: 0.547 / Mean I/σ(I) obs: 2.5 / Num. unique obs: 4794 / CC1/2: 0.787 / % possible all: 78.5

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Processing

Software
NameVersionClassification
REFMAC5.8.0238refinement
PHASER2.8.2phasing
STARANISOdata scaling
XDSversion Jan 2018data reduction
RefinementMethod to determine structure: MOLECULAR REPLACEMENT / Resolution: 1.8→43.46 Å / Cor.coef. Fo:Fc: 0.961 / Cor.coef. Fo:Fc free: 0.922 / SU B: 6.238 / SU ML: 0.093 / Cross valid method: THROUGHOUT / ESU R: 0.132 / ESU R Free: 0.135 / Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS
RfactorNum. reflection% reflectionSelection details
Rfree0.22209 1083 1.1 %RANDOM
Rwork0.16524 ---
obs0.16583 94875 87.89 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.2 Å
Displacement parametersBiso mean: 24.503 Å2
Baniso -1Baniso -2Baniso -3
1--0.21 Å20 Å2-0.16 Å2
2--0.5 Å2-0 Å2
3----0.19 Å2
Refinement stepCycle: LAST / Resolution: 1.8→43.46 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms8798 0 62 986 9846
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0130.0139161
X-RAY DIFFRACTIONr_bond_other_d0.0020.0178490
X-RAY DIFFRACTIONr_angle_refined_deg1.7961.63212448
X-RAY DIFFRACTIONr_angle_other_deg1.4631.57119813
X-RAY DIFFRACTIONr_dihedral_angle_1_deg7.81751159
X-RAY DIFFRACTIONr_dihedral_angle_2_deg34.27324.034414
X-RAY DIFFRACTIONr_dihedral_angle_3_deg14.21151498
X-RAY DIFFRACTIONr_dihedral_angle_4_deg9.6711528
X-RAY DIFFRACTIONr_chiral_restr0.0950.21184
X-RAY DIFFRACTIONr_gen_planes_refined0.0110.0210165
X-RAY DIFFRACTIONr_gen_planes_other0.0010.021800
X-RAY DIFFRACTIONr_nbd_refined
X-RAY DIFFRACTIONr_nbd_other
X-RAY DIFFRACTIONr_nbtor_refined
X-RAY DIFFRACTIONr_nbtor_other
X-RAY DIFFRACTIONr_xyhbond_nbd_refined
X-RAY DIFFRACTIONr_xyhbond_nbd_other
X-RAY DIFFRACTIONr_metal_ion_refined
X-RAY DIFFRACTIONr_metal_ion_other
X-RAY DIFFRACTIONr_symmetry_vdw_refined
X-RAY DIFFRACTIONr_symmetry_vdw_other
X-RAY DIFFRACTIONr_symmetry_hbond_refined
X-RAY DIFFRACTIONr_symmetry_hbond_other
X-RAY DIFFRACTIONr_symmetry_metal_ion_refined
X-RAY DIFFRACTIONr_symmetry_metal_ion_other
X-RAY DIFFRACTIONr_mcbond_it1.1421.0044587
X-RAY DIFFRACTIONr_mcbond_other1.1421.0034586
X-RAY DIFFRACTIONr_mcangle_it1.8551.4915726
X-RAY DIFFRACTIONr_mcangle_other1.8551.4915727
X-RAY DIFFRACTIONr_scbond_it1.4481.1644574
X-RAY DIFFRACTIONr_scbond_other1.4241.1584567
X-RAY DIFFRACTIONr_scangle_it
X-RAY DIFFRACTIONr_scangle_other2.2741.6526698
X-RAY DIFFRACTIONr_long_range_B_refined5.9813.30310533
X-RAY DIFFRACTIONr_long_range_B_other5.97913.30610534
X-RAY DIFFRACTIONr_rigid_bond_restr
X-RAY DIFFRACTIONr_sphericity_free
X-RAY DIFFRACTIONr_sphericity_bonded
LS refinement shellResolution: 1.799→1.846 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.255 47 -
Rwork0.231 3472 -
obs--43.94 %
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)
10.2939-0.13850.09130.10640.07080.40030.01030.0339-0.0184-0.0122-0.02280.0373-0.00840.06460.01250.2073-0.0113-0.00220.12820.00610.176119.24339.2111-11.9714
20.4995-0.2994-0.19920.20630.06461.58030.021-0.0003-0.0178-0.0397-0.01660.073-0.0339-0.075-0.00440.17790.01-0.01370.1298-0.00130.19541.016219.6404-1.1519
30.4474-0.2034-0.23371.54890.68670.3662-0.02610.1534-0.1899-0.0947-0.03590.0828-0.0389-0.09710.0620.16340.0143-0.0170.1506-0.00440.1921.670612.3805-13.019
40.9886-0.3516-0.0930.1423-0.03510.33130.0348-0.01650.1173-0.0228-0.0059-0.043-0.06510.0368-0.02880.2204-0.02240.00190.08580.00170.209914.149326.4911-0.4985
50.711-0.50190.3750.4732-0.31080.3867-0.0253-0.1862-0.087-0.00320.0646-0.0217-0.0244-0.1116-0.03930.15850.00070.01810.1663-0.01310.17336.36215.564711.1075
63.22131.29770.1120.543-0.07420.8975-0.0355-0.07970.01230.023-0.02260.0021-0.11570.02860.05810.1992-0.0097-0.01550.1357-0.01160.179622.575918.217712.2995
71.59570.9129-0.50190.5568-0.4481.06690.10260.0622-0.06820.0429-0.01870.00040.030.1614-0.08380.15720.03520.01210.1948-0.00820.168335.30812.9107-10.8676
80.3707-0.62570.2341.0888-0.36591.1640.00690.0531-0.0533-0.0008-0.06780.16080.01140.48740.06090.06120.03330.03210.29290.02310.181544.70020.8347-3.6394
90.5822-0.2010.31280.0716-0.13411.1118-0.0033-0.008-0.01270.0107-0.0145-0.0043-0.04120.14880.01790.1417-0.01980.00110.19150.01540.176635.485.36789.0951
100.7823-0.6746-0.3990.69570.5581.3459-0.0008-0.04990.01650.0299-0.0464-0.06070.04910.15010.04720.06620.0237-0.02680.26250.01960.16141.8990.524819.8544
111.71860.05421.35440.00560.05151.2047-0.04440.35980.09430.0129-0.0165-0.00140.030.50930.06090.05160.02950.00810.47310.0590.102450.52322.24866.5971
120.66090.087-0.30920.0173-0.04080.1471-0.0505-0.1893-0.0945-0.0316-0.011-0.0120.03550.11150.06150.12440.0711-0.01010.27340.06860.169842.7472-8.060819.388
130.3035-0.1190.2350.0561-0.12890.68440.0548-0.057-0.0586-0.05540.01640.03820.07580.0589-0.07120.18440.0131-0.01560.14740.03670.191827.5366-6.522812.4198
140.40580.2808-0.0281.1298-1.11041.5631-0.0409-0.15690.0962-0.0291-0.01480.02310.00220.10150.05560.12050.00140.00440.22510.01160.158330.01737.430522.7604
151.29330.77360.75850.51380.57840.8798-0.0213-0.1319-0.0268-0.0201-0.01740.0396-0.0040.00610.03870.1727-0.00410.00450.15260.03570.197417.62173.451713.4941
160.31980.0471-0.10020.01580.04280.678-0.0073-0.0178-0.08040.01-0.01270.0027-0.0530.08890.020.1751-0.01350.00970.1716-0.00030.16156.224110.737150.665
171.861.04640.14650.6151-0.06851.394-0.0202-0.2412-0.251-0.014-0.1848-0.1248-0.00390.2390.2050.09950.0375-0.01580.18330.02540.249415.6114.880854.9534
180.33360.04120.0010.01340.04830.69410.0436-0.0468-0.0713-0.0096-0.00420.0139-0.22680.1012-0.03940.2402-0.06950.01910.1463-0.02310.147411.317324.38544.2953
191.28010.91010.00140.70160.10090.83720.02340.0734-0.04360.00170.06330.0447-0.06540.2433-0.08680.1388-0.04080.0170.1993-0.02630.197613.446313.964630.9644
201.4796-1.9650.1253.00130.3270.636-0.04780.0448-0.0957-0.1720.05410.1091-0.29670.1579-0.00640.2249-0.02630.02390.1359-0.00220.15490.489322.29630.893
210.3966-0.3018-0.19911.17351.20512.16360.02560.0106-0.11260.0115-0.05230.0267-0.0683-0.1940.02670.16860.00610.01840.1703-0.02030.161-16.36878.485851.5723
221.65760.6381.0550.47780.09131.14110.0447-0.1165-0.01060.17070.05520.0214-0.1636-0.268-0.09990.15180.05770.06080.2302-0.02330.1455-26.347516.581447.0251
230.5732-0.1226-0.04220.0319-0.07691.3529-0.01040.05210.01770.00360.0042-0.0044-0.1139-0.21560.00620.17930.0360.01480.1717-0.01870.1612-17.023916.530632.8634
242.58891.066-1.06280.5856-0.24770.7464-0.03980.01630.0949-0.0985-0.00730.1053-0.0682-0.12220.04710.08590.066-0.01820.27750.02780.1146-24.822216.445721.8466
252.1997-1.60231.40011.6741-0.68121.1189-0.056-0.12070.0293-0.06560.0714-0.1216-0.1411-0.1247-0.01550.10340.07670.05090.2477-0.03120.1533-32.237219.982935.7731
260.24360.19620.05490.18940.06750.0663-0.02990.0493-0.02840.02370.0259-0.02810.0064-0.08930.0040.1220.0010.00860.2649-0.01310.1335-23.34344.837424.672
270.1636-0.0390.33020.3450.45961.6165-0.0177-0.0043-0.02590.020.0260.00280.0461-0.079-0.00830.1673-0.02050.01860.1663-0.0380.174-11.09273.809728.493
281.34330.320.14690.31040.76112.37140.05760.22470.0735-0.00190.0253-0.0497-0.1133-0.1576-0.08280.20740.0494-0.00660.149-0.01890.1614-10.567416.692919.2557
290.6734-0.8853-0.21382.9699-1.10271.1385-0.02620.07380.0013-0.028-0.0616-0.17120.0183-0.04170.08780.1534-0.01170.01430.1556-0.03560.1954-1.73717.2127.6686
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1A31 - 117
2X-RAY DIFFRACTION2A118 - 153
3X-RAY DIFFRACTION3A154 - 175
4X-RAY DIFFRACTION4A176 - 281
5X-RAY DIFFRACTION5A282 - 311
6X-RAY DIFFRACTION6A312 - 334
7X-RAY DIFFRACTION7B31 - 67
8X-RAY DIFFRACTION8B68 - 92
9X-RAY DIFFRACTION9B93 - 117
10X-RAY DIFFRACTION10B118 - 153
11X-RAY DIFFRACTION11B154 - 172
12X-RAY DIFFRACTION12B173 - 201
13X-RAY DIFFRACTION13B202 - 281
14X-RAY DIFFRACTION14B282 - 311
15X-RAY DIFFRACTION15B312 - 334
16X-RAY DIFFRACTION16C31 - 153
17X-RAY DIFFRACTION17C154 - 175
18X-RAY DIFFRACTION18C176 - 281
19X-RAY DIFFRACTION19C282 - 311
20X-RAY DIFFRACTION20C312 - 334
21X-RAY DIFFRACTION21D31 - 60
22X-RAY DIFFRACTION22D61 - 92
23X-RAY DIFFRACTION23D93 - 117
24X-RAY DIFFRACTION24D118 - 153
25X-RAY DIFFRACTION25D154 - 172
26X-RAY DIFFRACTION26D173 - 241
27X-RAY DIFFRACTION27D242 - 281
28X-RAY DIFFRACTION28D282 - 311
29X-RAY DIFFRACTION29D312 - 334

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