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- PDB-7kno: Crystal structure of Acetyl-CoA synthetase in complex with adenos... -

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Basic information

Entry
Database: PDB / ID: 7kno
TitleCrystal structure of Acetyl-CoA synthetase in complex with adenosine-5'-ethylphosphate from Cryptococcus neoformans var. grubii serotype A (H99)
ComponentsAcetyl-coenzyme A synthetase
KeywordsLIGASE / SSGCID / Structural Genomics / Seattle Structural Genomics Center for Infectious Disease
Function / homology
Function and homology information


acetate-CoA ligase / acetate-CoA ligase activity / acetyl-CoA biosynthetic process from acetate / AMP binding / mitochondrion / ATP binding / metal ion binding / cytosol
Similarity search - Function
Acetate-CoA ligase / Acetyl-coenzyme A synthetase, N-terminal domain / Acetyl-coenzyme A synthetase N-terminus / ANL, N-terminal domain / AMP-binding enzyme, C-terminal domain / AMP-binding enzyme C-terminal domain / AMP-binding, conserved site / Putative AMP-binding domain signature. / AMP-dependent synthetase/ligase / AMP-binding enzyme, C-terminal domain superfamily / AMP-binding enzyme
Similarity search - Domain/homology
PHOSPHATE ION / 5'-O-[(S)-ethoxy(hydroxy)phosphoryl]adenosine / Acetyl-coenzyme A synthetase
Similarity search - Component
Biological speciesCryptococcus neoformans var. grubii serotype A (fungus)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 1.8 Å
AuthorsSeattle Structural Genomics Center for Infectious Disease (SSGCID)
CitationJournal: to be published
Title: Crystal structure of Acetyl-CoA synthetase in complex with adenosine-5'-ethylphosphate from Cryptococcus neoformans var. grubii serotype A (H99)
Authors: Abendroth, J. / DeBouver, N.D. / Fox III, D. / Esan, T.E. / Hagen, T.J. / Krysan, D.J. / Lorimer, D.D. / Horanyi, P.S. / Edwards, T.E.
History
DepositionNov 5, 2020Deposition site: RCSB / Processing site: RCSB
Revision 1.0Dec 30, 2020Provider: repository / Type: Initial release
Revision 1.1Oct 18, 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: Acetyl-coenzyme A synthetase
B: Acetyl-coenzyme A synthetase
C: Acetyl-coenzyme A synthetase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)234,52419
Polymers232,4273
Non-polymers2,09716
Water31,2561735
1


  • Idetical with deposited unit
  • defined by author
  • Evidence: gel filtration
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area7500 Å2
ΔGint0 kcal/mol
Surface area63700 Å2
Unit cell
Length a, b, c (Å)72.690, 184.700, 84.840
Angle α, β, γ (deg.)90.000, 93.869, 90.000
Int Tables number4
Space group name H-MP1211
Space group name HallP2yb
Symmetry operation#1: x,y,z
#2: -x,y+1/2,-z

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Components

#1: Protein Acetyl-coenzyme A synthetase


Mass: 77475.750 Da / Num. of mol.: 3
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Cryptococcus neoformans var. grubii serotype A (strain H99 / ATCC 208821 / CBS 10515 / FGSC 9487) (fungus)
Strain: H99 / ATCC 208821 / CBS 10515 / FGSC 9487 / Gene: CNAG_00797 / Plasmid: Crnec.00629.a.FS11 / Production host: Escherichia coli BL21(DE3) (bacteria) / Strain (production host): BL21(DE3) / References: UniProt: J9VFT1, acetate-CoA ligase
#2: Chemical ChemComp-WTA / 5'-O-[(S)-ethoxy(hydroxy)phosphoryl]adenosine


Mass: 375.274 Da / Num. of mol.: 3 / Source method: obtained synthetically / Formula: C12H18N5O7P / Feature type: SUBJECT OF INVESTIGATION
#3: Chemical
ChemComp-EDO / 1,2-ETHANEDIOL / ETHYLENE GLYCOL


Mass: 62.068 Da / Num. of mol.: 8 / Source method: obtained synthetically / Formula: C2H6O2
#4: Chemical
ChemComp-PO4 / PHOSPHATE ION


Mass: 94.971 Da / Num. of mol.: 5 / Source method: obtained synthetically / Formula: PO4
#5: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 1735 / Source method: isolated from a natural source / Formula: H2O
Has ligand of interestY

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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: 2.45 Å3/Da / Density % sol: 49.8 %
Crystal growTemperature: 287 K / Method: vapor diffusion, sitting drop / pH: 6.2
Details: Optimization screen around RigakuReagents Wizard screen 1/2, condition e8: 100mM potassium / sodium phosphate dibasic pH 6.2, 200mM NaCl 13.18% (w/V) PEG 8000: CrneC.00629.a.FS11.PD00460 at ...Details: Optimization screen around RigakuReagents Wizard screen 1/2, condition e8: 100mM potassium / sodium phosphate dibasic pH 6.2, 200mM NaCl 13.18% (w/V) PEG 8000: CrneC.00629.a.FS11.PD00460 at 10mg/ml + ethyl-AMP + AcCoA: tray 317932 c7: cryo: 20% EG + compounds: puck: fgi6-9

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Data collection

DiffractionMean temperature: 100 K / Serial crystal experiment: N
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 21-ID-F / Wavelength: 0.97872 Å
DetectorType: RAYONIX MX-300 / Detector: CCD / Date: Sep 24, 2020 / Details: Beryllium Lenses
RadiationMonochromator: Diamond [111] / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.97872 Å / Relative weight: 1
ReflectionResolution: 1.8→50 Å / Num. obs: 205272 / % possible obs: 99.8 % / Redundancy: 3.78 % / Biso Wilson estimate: 32.568 Å2 / CC1/2: 0.999 / Rmerge(I) obs: 0.052 / Rrim(I) all: 0.06 / Χ2: 0.928 / Net I/σ(I): 15.12
Reflection shell

Diffraction-ID: 1

Resolution (Å)Redundancy (%)Rmerge(I) obsMean I/σ(I) obsNum. unique obsCC1/2Rrim(I) all% possible all
1.8-1.853.6270.5152.31151380.8490.60599.9
1.85-1.93.7940.4122.99148110.9020.4899.9
1.9-1.953.7970.323.89143420.9330.37399.9
1.95-2.013.8060.2624.77140230.9510.30599.9
2.01-2.083.8070.2066.05135470.9680.2499.9
2.08-2.153.8180.1637.5130450.9790.1999.9
2.15-2.233.8190.1269.6126430.9860.14699.9
2.23-2.323.8130.10611.28121650.9890.12499.8
2.32-2.433.8170.08713.43117030.9930.10299.8
2.43-2.553.8130.07415.57110970.9940.08699.9
2.55-2.683.8050.06318.14106530.9950.07399.9
2.68-2.853.7920.05321.0399980.9970.06299.8
2.85-3.043.7720.04424.4794510.9970.05299.8
3.04-3.293.7570.03828.8487840.9980.04499.8
3.29-3.63.7470.03332.8280510.9980.03899.7
3.6-4.023.7470.0335.7473370.9980.03599.7
4.02-4.653.7620.02938.2564370.9980.03499.8
4.65-5.693.7730.02938.7954860.9980.03399.9
5.69-8.053.7750.0338.8842320.9980.03599.8
8.05-503.5650.0340.5823290.9980.03698.1

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Processing

Software
NameVersionClassification
PHENIX1.19rc4 4035refinement
XDSdata reduction
XSCALEdata scaling
PDB_EXTRACT3.27data extraction
MoRDaphasing
PHASERphasing
PHENIXmodel building
Cootmodel building
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: pdb entry 5ifi
Resolution: 1.8→46.94 Å / SU ML: 0.2077 / Cross valid method: FREE R-VALUE / σ(F): 1.35 / Phase error: 20.426
Stereochemistry target values: GeoStd + Monomer Library + CDL v1.2
RfactorNum. reflection% reflectionSelection details
Rfree0.1898 2070 1.01 %0
Rwork0.1637 203120 --
obs0.164 205190 99.84 %-
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL
Displacement parametersBiso mean: 37.97 Å2
Refinement stepCycle: LAST / Resolution: 1.8→46.94 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms14797 0 132 1735 16664
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.008215549
X-RAY DIFFRACTIONf_angle_d0.914121254
X-RAY DIFFRACTIONf_chiral_restr0.06152307
X-RAY DIFFRACTIONf_plane_restr0.00952752
X-RAY DIFFRACTIONf_dihedral_angle_d12.81855590
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
1.8-1.840.29251570.271113466X-RAY DIFFRACTION99.79
1.84-1.890.25321480.234113511X-RAY DIFFRACTION99.88
1.89-1.940.22691540.205413445X-RAY DIFFRACTION99.9
1.94-20.21871670.190413538X-RAY DIFFRACTION99.9
2-2.060.21981310.186313556X-RAY DIFFRACTION99.94
2.06-2.130.23041320.187313563X-RAY DIFFRACTION99.92
2.13-2.220.23371430.173913543X-RAY DIFFRACTION99.95
2.22-2.320.20861020.1713492X-RAY DIFFRACTION99.84
2.32-2.440.21321200.170413589X-RAY DIFFRACTION99.88
2.44-2.60.22841230.168613567X-RAY DIFFRACTION99.87
2.6-2.80.18781190.170913561X-RAY DIFFRACTION99.88
2.8-3.080.24581140.168613541X-RAY DIFFRACTION99.81
3.08-3.520.171430.154813559X-RAY DIFFRACTION99.7
3.52-4.440.15861540.134113570X-RAY DIFFRACTION99.75
4.44-46.940.14831630.145213619X-RAY DIFFRACTION99.57
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)
10.937373726407-0.4780786240410.2484994620490.863713732196-0.5591286488710.6307726542970.0255159488731-0.0367665856203-0.107398282081-0.1302445617290.205818436720.4377703166440.11921968727-0.235870150005-0.1458066114120.154181119463-0.0352667576894-0.0274156743630.2750241195050.07824747555270.294296311016-20.67853374773.16495980867-8.38998995232
20.938267073646-0.6548015015390.4652347293081.64215770985-0.8794047196451.069260953810.1835711513930.167008101029-0.159217955768-0.5005096353430.01108847562260.2238291884350.3901676343420.0282593219546-0.1301360824140.269907123675-0.00864277447757-0.04697124768810.206184475272-0.02165813633440.174176317881-8.48033983511-7.67361428003-17.4011025237
30.9478986052010.2611967844730.2884102114942.017501258-0.3327735749141.48971821319-0.01308189772860.02924512414910.06077002632-0.2640584535110.08505509905530.263329395044-0.0276075729337-0.115562542768-0.06694505302440.136336724958-0.00708555386060.00188355633750.1984943642690.03658999430830.197930452211-17.384115008917.2870503027-22.7072513962
42.128179118470.6090880443240.6435365948731.067926542871.741626743592.913991887830.02051791278810.484435437152-0.105523626179-0.9214509602010.2118926873670.2319258780580.0923949915699-0.0795276273815-0.2008185385680.792265838956-0.0698623509175-0.2703963839230.4200171367390.0338836394760.383765346994-22.79400487085.69955693627-46.8315315069
55.80420438142-0.5229508102683.547220442083.36201089592-0.9730414269472.775087787980.03856398911940.470591374196-0.345609283599-0.651460952874-0.050085310559-0.4054400564480.8325284143170.521874063156-0.06700974924440.6872719645370.05528473847870.01557597190360.506283201344-0.01871339716710.351039784544-6.872322822252.0371233721-40.8033519266
60.654225168357-0.3474852619380.3560199615141.88382635094-0.6097173228460.690725290923-0.145468630918-0.1036523855660.01086612359120.4736748752780.147808412814-0.0306055418713-0.127148643993-0.07633564748213.23205696739E-50.2571605557320.06656126134150.01715611662460.204194076793-0.02620572925210.099180074093512.7871903037-20.426764238923.3644557837
72.014850190730.1898419747250.1835523869394.03197768966-0.3699088612942.0783688594-0.08721239497150.116235067850.008766858970170.007416111032650.0783104533099-0.768618577437-0.2312903686680.2609648391290.03456425889750.224291140397-0.001750824406180.02879400793770.22160971404-0.04235630244930.30847891051323.2607467533-47.565105031123.566201342
80.8601360445080.04021034516490.278216094350.2206085906130.2432936204250.47411763568-0.447700199971-0.3315775699970.6348837886680.7232013891990.0677144817738-0.276106083281-0.656300206534-0.1763722222380.1505579524350.7504486571940.163798445242-0.2708064338870.304415169139-0.1861071856080.4699100624555.3679814385926.642071675118.8123083548
91.01265433390.06580398974020.106028991450.140457737807-0.1421770507340.471405696753-0.2982486182790.2579919300590.5858232578770.241025349069-0.194045459572-0.500867924696-0.389003596350.2661310624210.2200436331770.184206085161-0.101320257041-0.1180613950060.3228478927630.1967149886660.59290151367918.548323007921.8785547119-5.23188378292
100.2760737329950.04920405132220.08747884044971.29634667057-0.2482311794490.722178649834-0.410430507942-0.1070693340230.685063739740.420472481673-0.08926779105-0.343344596031-0.7761506966040.1560008220230.07672154016730.680533654892-0.0316363827536-0.4127669359630.152969439823-0.03326204663810.71185429990813.811903925232.639306443510.2387009407
110.6190331899890.4254364168350.2078903269220.3607367467550.1797753828980.0826998082772-0.167665267426-0.03681881811590.5050393414280.145287559524-0.0399977537117-0.155483725974-0.5584299438880.1102238434870.232985021311.28544996333-0.0963615620152-0.5888400475920.28047448198-0.1379063755251.0745957900216.84074519449.830348415515.5008482688
120.5130536628380.1895089165360.08962201788270.8209311214110.293466033470.709763743767-0.285405604171-0.5456085415260.530139954720.468196989414-0.102519518712-0.360805266323-0.6395918011670.274978531784-0.5398050297290.960716942499-0.0516921743726-0.7763619124660.324420742615-0.3094227158380.65173385445223.035960088627.979919852527.5248851054
131.433644718331.63252203698-1.242596942014.13769987343-0.569704920253.58720370784-0.0333549799770.03527885043880.559446733957-0.07812586268370.078031510988-0.242221359305-0.09695285152340.491186668708-0.04984887013290.752747751599-0.0349693873053-0.01915924961990.934338280214-0.03350549998530.9842596345446.392919226731.921093814910.5386016021
140.0736919000910.4754556469740.2476609703524.096396439450.04062123353935.5664680726-0.2091499579360.3254450079580.765183339702-0.233919449774-0.02103485986240.0188654015232-0.6043819223520.8773483707120.2104808804310.639403227655-0.0766959572084-0.07967603520210.8028649776320.001510913401651.0419222833144.525969748936.94320999656.04303682549
Refinement TLS group

Refine-ID: X-RAY DIFFRACTION

IDRefine TLS-IDSelection detailsAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
11chain 'A' and (resid 10 through 132 )AA10 - 1321 - 123
22chain 'A' and (resid 133 through 453 )AA133 - 453124 - 442
33chain 'A' and (resid 454 through 555 )AA454 - 555443 - 544
44chain 'A' and (resid 556 through 629 )AA556 - 629545 - 618
55chain 'A' and (resid 630 through 677 )AA630 - 677619 - 666
66chain 'B' and (resid 10 through 555 )BG10 - 5551 - 542
77chain 'B' and (resid 556 through 677 )BG556 - 677543 - 664
88chain 'C' and (resid 25 through 132 )CN25 - 1321 - 108
99chain 'C' and (resid 133 through 278 )CN133 - 278109 - 254
1010chain 'C' and (resid 279 through 382 )CN279 - 382255 - 358
1111chain 'C' and (resid 383 through 432 )CN383 - 432359 - 408
1212chain 'C' and (resid 433 through 551 )CN433 - 551409 - 527
1313chain 'C' and (resid 552 through 602 )CN552 - 602528 - 564
1414chain 'C' and (resid 603 through 633 )CN603 - 633565 - 595

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