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- PDB-5f7j: Crystal structure of Mutant N87T of adenosine/Methylthioadenosine... -

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Basic information

Entry
Database: PDB / ID: 5f7j
TitleCrystal structure of Mutant N87T of adenosine/Methylthioadenosine phosphorylase from Schistosoma mansoni in complex with Adenine
ComponentsMethylthioadenosine phosphorylase
KeywordsTRANSFERASE
Function / homology
Function and homology information


S-methyl-5'-thioadenosine phosphorylase / S-methyl-5-thioadenosine phosphorylase activity / L-methionine salvage from methylthioadenosine / purine ribonucleoside salvage / nucleus / cytosol
Similarity search - Function
Methylthioadenosine phosphorylase (MTAP) / Purine phosphorylase, family 2, conserved site / Purine and other phosphorylases family 2 signature. / Nucleoside phosphorylase domain / Nucleoside phosphorylase domain / Phosphorylase superfamily / Nucleoside phosphorylase superfamily / Rossmann fold / 3-Layer(aba) Sandwich / Alpha Beta
Similarity search - Domain/homology
ADENINE / PHOSPHATE ION / S-methyl-5'-thioadenosine phosphorylase
Similarity search - Component
Biological speciesSchistosoma mansoni (invertebrata)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / molecular replacement / Resolution: 1.66 Å
AuthorsTorini, J.R. / Brandao-Neto, J. / DeMarco, R. / Pereira, H.M.
Funding support Brazil, 2items
OrganizationGrant numberCountry
Sao Paulo Research Foundation (FAPESP)2012/14223-9 Brazil
Brazilian National Council for Scientific and Technological Development (CNPq)474402/2013-4 Brazil
CitationJournal: PLoS Negl Trop Dis / Year: 2016
Title: Crystal Structure of Schistosoma mansoni Adenosine Phosphorylase/5'-Methylthioadenosine Phosphorylase and Its Importance on Adenosine Salvage Pathway.
Authors: Torini, J.R. / Brandao-Neto, J. / DeMarco, R. / Pereira, H.D.
History
DepositionDec 8, 2015Deposition site: RCSB / Processing site: RCSB
Revision 1.0Dec 14, 2016Provider: repository / Type: Initial release
Revision 1.1Dec 21, 2016Group: Database references
Revision 1.2Sep 27, 2017Group: Author supporting evidence / Data collection / Category: diffrn_source / pdbx_audit_support
Item: _diffrn_source.pdbx_synchrotron_site / _pdbx_audit_support.funding_organization
Revision 1.3Apr 17, 2019Group: Author supporting evidence / Data collection / Category: pdbx_audit_support / Item: _pdbx_audit_support.funding_organization
Revision 1.4Jan 1, 2020Group: Author supporting evidence / Category: pdbx_audit_support / Item: _pdbx_audit_support.funding_organization
Revision 1.5Sep 27, 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: Methylthioadenosine phosphorylase
B: Methylthioadenosine phosphorylase
C: Methylthioadenosine phosphorylase
D: Methylthioadenosine phosphorylase
E: Methylthioadenosine phosphorylase
F: Methylthioadenosine phosphorylase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)212,39914
Polymers211,4796
Non-polymers9208
Water31,1301728
1
A: Methylthioadenosine phosphorylase
B: Methylthioadenosine phosphorylase
C: Methylthioadenosine phosphorylase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)106,2007
Polymers105,7393
Non-polymers4604
Water543
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area8390 Å2
ΔGint-56 kcal/mol
Surface area32180 Å2
MethodPISA
2
D: Methylthioadenosine phosphorylase
E: Methylthioadenosine phosphorylase
F: Methylthioadenosine phosphorylase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)106,2007
Polymers105,7393
Non-polymers4604
Water543
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area8270 Å2
ΔGint-55 kcal/mol
Surface area31250 Å2
MethodPISA
Unit cell
Length a, b, c (Å)81.493, 82.106, 150.267
Angle α, β, γ (deg.)90.00, 101.10, 90.00
Int Tables number4
Space group name H-MP1211

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Components

#1: Protein
Methylthioadenosine phosphorylase


Mass: 35246.484 Da / Num. of mol.: 6 / Fragment: Enzyme / Mutation: N87T
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Schistosoma mansoni (invertebrata) / Plasmid: pET28a / Production host: Escherichia coli (E. coli) / Strain (production host): BL21(DE3)
References: UniProt: I0B503, S-methyl-5'-thioadenosine phosphorylase
#2: Chemical
ChemComp-ADE / ADENINE


Mass: 135.127 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: C5H5N5
#3: Chemical
ChemComp-PO4 / PHOSPHATE ION


Mass: 94.971 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: PO4
#4: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 1728 / 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: 2.33 Å3/Da / Density % sol: 47.25 %
Crystal growTemperature: 291 K / Method: vapor diffusion, hanging drop / pH: 6.5 / Details: 100mM Bris-tris or Mes, 14-18% PEG3350, pH 6.5 / PH range: 6.1-6.7

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: Diamond / Beamline: I04 / Wavelength: 0.97949 Å
DetectorType: DECTRIS PILATUS 6M-F / Detector: PIXEL / Date: Jul 21, 2015
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.97949 Å / Relative weight: 1
ReflectionResolution: 1.66→76.76 Å / Num. obs: 221276 / % possible obs: 96.5 % / Redundancy: 3.8 % / Biso Wilson estimate: 19.42 Å2 / Rmerge(I) obs: 0.084 / Net I/σ(I): 7.8 / Scaling rejects: 1153
Reflection shell
Resolution (Å)Redundancy (%)Rmerge(I) obsDiffraction-IDRejects% possible all
1.66-1.722.30.7371075.7
6.21-76.764.10.0491099.9

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Phasing

PhasingMethod: molecular replacement

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Processing

Software
NameVersionClassification
PHENIX(1.10.1_2155: ???)refinement
xia2data scaling
PHASERphasing
PDB_EXTRACT3.15data extraction
xia2data reduction
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 4L6I
Resolution: 1.66→73.727 Å / SU ML: 0.23 / Cross valid method: FREE R-VALUE / σ(F): 1.33 / Phase error: 28 / Stereochemistry target values: ML
RfactorNum. reflection% reflectionSelection details
Rfree0.2263 10728 4.91 %Random selection
Rwork0.1946 ---
obs0.1961 218591 95.34 %-
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL
Refinement stepCycle: LAST / Resolution: 1.66→73.727 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms13034 0 60 1728 14822
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.00813404
X-RAY DIFFRACTIONf_angle_d0.89518187
X-RAY DIFFRACTIONf_dihedral_angle_d14.7618041
X-RAY DIFFRACTIONf_chiral_restr0.0592105
X-RAY DIFFRACTIONf_plane_restr0.0062326
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
1.66-1.67890.38422460.38334921X-RAY DIFFRACTION68
1.6789-1.69860.40772720.37215267X-RAY DIFFRACTION73
1.6986-1.71930.40623000.36765550X-RAY DIFFRACTION78
1.7193-1.74110.39233110.35966023X-RAY DIFFRACTION83
1.7411-1.7640.40113410.34076379X-RAY DIFFRACTION88
1.764-1.78820.34983510.32376767X-RAY DIFFRACTION93
1.7882-1.81370.343960.29957057X-RAY DIFFRACTION98
1.8137-1.84080.35923850.28957155X-RAY DIFFRACTION99
1.8408-1.86960.30383620.28667168X-RAY DIFFRACTION99
1.8696-1.90020.31823820.30647142X-RAY DIFFRACTION98
1.9002-1.9330.40713330.35786748X-RAY DIFFRACTION94
1.933-1.96820.29813350.24737164X-RAY DIFFRACTION99
1.9682-2.0060.25213890.21377203X-RAY DIFFRACTION100
2.006-2.0470.23323670.21577239X-RAY DIFFRACTION100
2.047-2.09150.31423750.24227073X-RAY DIFFRACTION98
2.0915-2.14010.23333430.1957272X-RAY DIFFRACTION100
2.1401-2.19370.24343900.19167190X-RAY DIFFRACTION100
2.1937-2.2530.25143630.21527150X-RAY DIFFRACTION98
2.253-2.31930.23233520.18867186X-RAY DIFFRACTION98
2.3193-2.39410.21063670.1667239X-RAY DIFFRACTION100
2.3941-2.47970.21193590.177253X-RAY DIFFRACTION100
2.4797-2.5790.19763770.16937256X-RAY DIFFRACTION100
2.579-2.69640.22273570.17487272X-RAY DIFFRACTION100
2.6964-2.83850.20583820.16547273X-RAY DIFFRACTION100
2.8385-3.01640.21943800.1697287X-RAY DIFFRACTION100
3.0164-3.24930.19844060.16487249X-RAY DIFFRACTION100
3.2493-3.57630.19713630.16917328X-RAY DIFFRACTION100
3.5763-4.09370.17243630.15297334X-RAY DIFFRACTION100
4.0937-5.15750.15313620.13637348X-RAY DIFFRACTION100
5.1575-73.80.18034190.17887370X-RAY DIFFRACTION99
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)
12.11760.4128-0.32781.65-0.72531.47860.0099-0.0571-0.18160.12780.00310.3380.1683-0.27340.01910.1559-0.03980.03720.16970.00540.2489-11.6523-21.697369.775
21.71360.3332-0.16691.07550.16371.31010.04440.10810.0778-0.0557-0.03490.1437-0.0862-0.20740.00420.120.04130.00450.12870.02110.1248-5.6485-6.943959.9535
37.88092.56314.42821.5372.66434.81680.2407-1.08430.34440.2489-0.29030.2286-0.1124-0.60910.08650.2590.06510.07160.22160.0330.2781-9.1426-3.172173.5386
43.4156-0.70410.28011.5281-0.95070.60610.05970.6998-0.7011-0.4415-0.21880.39560.3966-0.52220.14610.2451-0.1018-0.12220.4387-0.09070.4066-16.5112-23.789349.2505
50.9063-1.0942-0.65478.83211.76034.7776-0.32790.6235-0.8477-0.3734-0.2790.77320.5661-0.45990.48690.3073-0.0260.00930.3024-0.09830.271915.522-33.823730.4705
61.1954-0.351-0.34510.92290.39141.3120.03640.1815-0.0565-0.1038-0.05990.0474-0.0378-0.13730.02190.1850.01780.01610.1462-0.00710.11113.9552-19.779738.1809
70.9106-0.0784-0.49051.34480.74283.18590.10310.3447-0.1708-0.1949-0.12790.1622-0.0999-0.29230.01690.27350.04120.03580.130.02810.169213.383-19.977733.3852
82.81270.49814.50341.7706-0.62268.51470.22440.2755-0.18890.0747-0.1074-0.64220.56520.987-0.13780.19140.0083-0.0180.20990.02290.308238.4638-9.972872.8868
97.71811.8718-0.41573.0395-1.43111.4495-0.0402-0.4602-0.28750.2241-0.0974-0.37060.08810.38480.12510.20710.035-0.01130.17950.01830.211330.6324-22.018675.7218
100.86740.0595-0.22931.5261-0.2570.70560.0549-0.01710.0810.0022-0.0193-0.0703-0.0780.0633-0.03910.141-0.00540.00360.1028-0.00720.114225.8532-4.417667.4584
110.73780.14050.39811.3365-0.20651.96110.05090.11730.0118-0.16850.0169-0.0466-0.09910.0288-0.05720.14920.00920.02640.1025-0.00690.139322.8165-5.7457.8464
122.8384-0.2045-2.43062.17030.97992.95790.05010.0540.0776-0.24720.1003-0.4956-0.0140.4542-0.12620.1937-0.03050.03680.12470.02820.255140.2417-1.301463.0718
136.35562.0519-0.78123.2708-0.37182.53020.1015-0.52570.02030.2783-0.07460.31660.0503-0.3644-0.04180.28790.01910.06270.20310.02960.161711.7141-5.127484.1399
144.5324-2.299-1.36613.8296-2.33123.85720.1183-0.20020.21380.43590.21780.6564-0.0519-0.5427-0.20830.1826-0.04270.06620.30110.03070.414-3.6956-22.1281-0.8958
152.04690.5038-0.35861.9457-0.56091.67030.0153-0.1158-0.35510.2002-0.02680.23160.2353-0.25660.03690.1777-0.03610.02310.18230.0350.25575.461-23.853-4.7244
161.81610.0516-0.0521.1834-0.09851.58450.0150.0841-0.0294-0.10920.00140.0963-0.1419-0.2187-0.00780.12170.041-0.00930.14850.01830.11137.9367-8.4948-14.1367
177.33860.0610.34391.9747-2.846.91680.0793-0.68922.04240.34030.1854-0.166-2.16190.1957-0.18170.6491-0.03860.0520.3024-0.08080.590819.58624.7518-1.0082
183.43780.4005-0.05932.81380.84442.44810.0781-0.0836-0.24890.0319-0.09910.37320.0595-0.57930.08120.16640.0246-0.04350.2060.13530.2332-0.7837-15.9153-10.9743
190.5092-1.0796-0.39462.53741.74076.6989-0.23280.2043-0.63540.0747-0.06640.58270.564-0.37130.37250.36090.01960.14620.2728-0.05890.49429.5299-34.9949-44.0544
201.3381-0.4705-0.72871.18040.54851.8018-0.01560.0555-0.1309-0.1353-0.0331-0.0673-0.0367-0.01540.0480.18940.00620.05070.1711-0.02710.179627.4333-20.5728-36.5692
211.0454-0.0069-0.42041.78461.07664.2419-0.03220.2209-0.2147-0.2358-0.150.1485-0.0488-0.38790.13760.32290.02810.07460.1420.01130.267826.4362-21.2143-41.5177
221.0606-1.002-0.44712.86950.99081.93360.2003-0.4171-0.32350.226-0.1284-0.42280.25550.4296-0.00330.1833-0.0724-0.03990.44330.18460.346848.3532-10.0003-1.9797
231.8280.2122-0.48041.6026-0.49850.33860.0755-0.7117-0.49990.394-0.2951-0.29250.04270.51360.16010.2854-0.0297-0.05120.56260.24570.420848.0341-15.29731.6568
241.70180.31570.06592.0131-0.37462.0854-0.0317-0.3351-0.42640.0704-0.2324-0.26570.2150.36510.2180.1621-0.00030.02250.25860.11890.303542.0185-13.8745-6.0059
252.46340.7633-0.861.4179-0.32411.57390.155-0.11440.06930.0295-0.0889-0.1002-0.20730.1749-0.06280.1576-0.02040.00560.15310.01710.14736.8289-0.2278-9.9817
263.7691-0.4148-1.00434.00540.07453.7624-0.0770.36970.0961-0.3556-0.1197-0.3338-0.25280.03830.15970.1993-0.02990.01380.1905-0.02360.165832.8077-7.5214-27.1282
272.39680.9537-0.58042.427-1.17394.3208-0.01850-0.131-0.2285-0.1002-0.2184-0.08590.24070.08870.1548-0.00640.02690.13620.02390.184737.6675-5.0856-13.2933
284.49060.1089-3.33143.08071.21983.57220.1472-0.36540.0179-0.2055-0.0775-0.3337-0.15650.6425-0.06870.2097-0.0544-0.01030.25870.11290.289154.0147-0.6429-11.3401
296.09622.8193-0.05953.01920.07212.14820.4617-0.99190.22750.5259-0.43690.1128-0.31770.1141-0.03980.3629-0.09040.02210.36790.01680.150829.7004-4.45158.9523
300.4411-0.1307-1.80310.00310.47726.6646-0.27410.1358-0.14790.0596-0.07970.03211.1722-0.20970.38030.35050.03050.03230.3108-0.02750.21658.3454-11.553517.5427
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection details
1X-RAY DIFFRACTION1chain 'A' and (resid 3 through 90 )
2X-RAY DIFFRACTION2chain 'A' and (resid 91 through 235 )
3X-RAY DIFFRACTION3chain 'A' and (resid 236 through 269 )
4X-RAY DIFFRACTION4chain 'A' and (resid 270 through 292 )
5X-RAY DIFFRACTION5chain 'B' and (resid 1 through 26 )
6X-RAY DIFFRACTION6chain 'B' and (resid 27 through 230 )
7X-RAY DIFFRACTION7chain 'B' and (resid 231 through 291 )
8X-RAY DIFFRACTION8chain 'C' and (resid 3 through 23 )
9X-RAY DIFFRACTION9chain 'C' and (resid 24 through 38 )
10X-RAY DIFFRACTION10chain 'C' and (resid 39 through 182 )
11X-RAY DIFFRACTION11chain 'C' and (resid 183 through 242 )
12X-RAY DIFFRACTION12chain 'C' and (resid 243 through 269 )
13X-RAY DIFFRACTION13chain 'C' and (resid 270 through 299 )
14X-RAY DIFFRACTION14chain 'D' and (resid 3 through 26 )
15X-RAY DIFFRACTION15chain 'D' and (resid 27 through 90 )
16X-RAY DIFFRACTION16chain 'D' and (resid 91 through 235 )
17X-RAY DIFFRACTION17chain 'D' and (resid 236 through 242 )
18X-RAY DIFFRACTION18chain 'D' and (resid 243 through 291 )
19X-RAY DIFFRACTION19chain 'E' and (resid 1 through 26 )
20X-RAY DIFFRACTION20chain 'E' and (resid 27 through 230 )
21X-RAY DIFFRACTION21chain 'E' and (resid 231 through 291 )
22X-RAY DIFFRACTION22chain 'F' and (resid 3 through 27 )
23X-RAY DIFFRACTION23chain 'F' and (resid 28 through 53 )
24X-RAY DIFFRACTION24chain 'F' and (resid 54 through 91 )
25X-RAY DIFFRACTION25chain 'F' and (resid 92 through 182 )
26X-RAY DIFFRACTION26chain 'F' and (resid 183 through 198 )
27X-RAY DIFFRACTION27chain 'F' and (resid 199 through 243 )
28X-RAY DIFFRACTION28chain 'F' and (resid 244 through 269 )
29X-RAY DIFFRACTION29chain 'F' and (resid 270 through 293 )
30X-RAY DIFFRACTION30(chain 'A' and resid 301) or (chain 'B' and resid 301) or (chain 'D' and resid 301) or (chain 'E' and resid 301)

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