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- PDB-5vn2: Crystal structure of 3-oxoacyl-[acyl-carrier protein] reductase f... -

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Basic information

Entry
Database: PDB / ID: 5vn2
TitleCrystal structure of 3-oxoacyl-[acyl-carrier protein] reductase from Brucella melitensis in complex with NAD
Components3-oxoacyl-(Acyl-carrier protein) reductase
KeywordsOXIDOREDUCTASE / SDR / short chain dehydrogenase reductase / Structural Genomics / Seattle Structural Genomics Center for Infectious Disease / SSGCID
Function / homology
Function and homology information


3-oxoacyl-[acyl-carrier-protein] reductase / 3-oxoacyl-[acyl-carrier-protein] reductase (NADPH) activity
Similarity search - Function
Short-chain dehydrogenase/reductase, conserved site / Short-chain dehydrogenases/reductases family signature. / Enoyl-(Acyl carrier protein) reductase / Short-chain dehydrogenase/reductase SDR / NAD(P)-binding Rossmann-like Domain / NAD(P)-binding domain superfamily / Rossmann fold / 3-Layer(aba) Sandwich / Alpha Beta
Similarity search - Domain/homology
ACETATE ION / NICOTINAMIDE-ADENINE-DINUCLEOTIDE / 3-oxoacyl-(Acyl-carrier protein) reductase
Similarity search - Component
Biological speciesBrucella melitensis biotype 1 (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / molecular replacement / Resolution: 1.9 Å
AuthorsSeattle Structural Genomics Center for Infectious Disease (SSGCID)
CitationJournal: to be published
Title: Crystal structure of 3-oxoacyl-[acyl-carrier protein] reductase from Brucella melitensis in complex with NAD
Authors: Mayclin, S.J. / Abendroth, J. / Lorimer, D.D. / Edwards, T.E.
History
DepositionApr 28, 2017Deposition site: RCSB / Processing site: RCSB
Revision 1.0May 31, 2017Provider: repository / Type: Initial release
Revision 1.1Oct 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: 3-oxoacyl-(Acyl-carrier protein) reductase
B: 3-oxoacyl-(Acyl-carrier protein) reductase
C: 3-oxoacyl-(Acyl-carrier protein) reductase
D: 3-oxoacyl-(Acyl-carrier protein) reductase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)113,88413
Polymers110,9324
Non-polymers2,9529
Water14,106783
1


  • Idetical with deposited unit
  • defined by author&software
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area19970 Å2
ΔGint-152 kcal/mol
Surface area33100 Å2
MethodPISA
Unit cell
Length a, b, c (Å)74.560, 92.710, 129.910
Angle α, β, γ (deg.)90.000, 90.000, 90.000
Int Tables number19
Space group name H-MP212121

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Components

#1: Protein
3-oxoacyl-(Acyl-carrier protein) reductase


Mass: 27733.045 Da / Num. of mol.: 4 / Fragment: residues 47-300
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Brucella melitensis biotype 1 (strain 16M / ATCC 23456 / NCTC 10094) (bacteria)
Strain: 16M / ATCC 23456 / NCTC 10094 / Gene: BMEI0026 / Production host: Escherichia coli (E. coli) / Strain (production host): BL21(DE3)
References: UniProt: Q8YJQ6, 3-oxoacyl-[acyl-carrier-protein] reductase
#2: Chemical
ChemComp-NAD / NICOTINAMIDE-ADENINE-DINUCLEOTIDE / Nicotinamide adenine dinucleotide


Mass: 663.425 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: C21H27N7O14P2 / Comment: NAD*YM
#3: Chemical
ChemComp-ACT / ACETATE ION / Acetate


Mass: 59.044 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: C2H3O2
#4: Chemical ChemComp-EDO / 1,2-ETHANEDIOL / ETHYLENE GLYCOL / Ethylene glycol


Mass: 62.068 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C2H6O2
#5: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 783 / 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: 2.03 Å3/Da / Density % sol: 39.3 %
Crystal growTemperature: 290 K / Method: vapor diffusion, sitting drop / pH: 7.5
Details: MCSG1 F5 (273386f5): 20% PEG3350, 20mM Sodium acetate, 4mM NAD, protein conc. 19.9mg/mL, cryo 15% ethylene glycol: unique puck ID: jif2-4

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 21-ID-F / Wavelength: 0.97872 Å
DetectorType: RAYONIX MX-300 / Detector: CCD / Date: Jun 22, 2016
RadiationMonochromator: Diamond [111] / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.97872 Å / Relative weight: 1
ReflectionResolution: 1.9→50 Å / Num. obs: 71305 / % possible obs: 99.6 % / Observed criterion σ(I): -3 / Redundancy: 5.407 % / Biso Wilson estimate: 21 Å2 / CC1/2: 0.999 / Rmerge(I) obs: 0.073 / Rrim(I) all: 0.081 / Χ2: 0.976 / Net I/σ(I): 17.22
Reflection shell

Diffraction-ID: 1

Resolution (Å)Redundancy (%)Rmerge(I) obsMean I/σ(I) obsNum. unique obsCC1/2Rrim(I) all% possible all
1.9-1.955.4070.5572.9952320.8580.61799.9
1.95-25.4140.4413.7450890.9050.48999.9
2-2.065.4040.3564.649690.9360.39499.9
2.06-2.125.4350.285.7548280.9580.31100
2.12-2.195.4180.2267.0246630.9720.25199.9
2.19-2.275.4180.2017.8345330.9770.22399.8
2.27-2.365.4370.1589.8243560.9860.17599.8
2.36-2.455.4330.13811.242090.9890.15299.8
2.45-2.565.4360.11813.0440510.9910.1399.8
2.56-2.695.4370.10414.7438450.9930.11599.8
2.69-2.835.430.08717.2637260.9950.09699.7
2.83-35.4250.06921.2534750.9960.07699.7
3-3.215.4340.05625.7332820.9980.06199.5
3.21-3.475.4210.04331.9530690.9980.04899.4
3.47-3.85.4130.03438.4128240.9990.03899.3
3.8-4.255.4060.02944.7325590.9990.03298.7
4.25-4.915.380.02550.0922720.9990.02898.8
4.91-6.015.3140.02647.1519470.9990.02998.6
6.01-8.55.2350.02550.1515180.9990.02797.6
8.5-504.7830.0257.8485810.02294.3

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Phasing

PhasingMethod: molecular replacement

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Processing

Software
NameVersionClassification
XSCALEdata scaling
Cootmodel building
PHENIX(dev_2744)refinement
PDB_EXTRACT3.22data extraction
XDSdata reduction
MOLREPphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 4one
Resolution: 1.9→50 Å / SU ML: 0.19 / Cross valid method: FREE R-VALUE / σ(F): 1.34 / Phase error: 19.42
RfactorNum. reflection% reflection
Rfree0.1983 1965 2.76 %
Rwork0.1499 --
obs0.1512 71283 99.52 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å
Displacement parametersBiso max: 97.32 Å2 / Biso mean: 29.2548 Å2 / Biso min: 9.24 Å2
Refinement stepCycle: final / Resolution: 1.9→50 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms7388 0 196 793 8377
Biso mean--23.97 35.55 -
Num. residues----1018
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.0067814
X-RAY DIFFRACTIONf_angle_d0.77310619
X-RAY DIFFRACTIONf_chiral_restr0.0541258
X-RAY DIFFRACTIONf_plane_restr0.0051446
X-RAY DIFFRACTIONf_dihedral_angle_d14.2744679
LS refinement shell

Refine-ID: X-RAY DIFFRACTION / Rfactor Rfree error: 0 / Total num. of bins used: 14

Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkNum. reflection all% reflection obs (%)
1.9-1.94750.27981540.233248775031100
1.9475-2.00010.25161470.211949095056100
2.0001-2.0590.25471470.194148895036100
2.059-2.12550.22221440.174749225066100
2.1255-2.20140.19891360.164749305066100
2.2014-2.28960.22041340.161749155049100
2.2896-2.39380.21711240.153949245048100
2.3938-2.520.21051520.154749165068100
2.52-2.67780.21261570.153549385095100
2.6778-2.88460.20131410.150449715112100
2.8846-3.17480.20611470.150249475094100
3.1748-3.63410.16821250.13385003512899
3.6341-4.5780.17811250.11875025515099
4.578-48.99250.15311320.13755152528497
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)
13.7766-0.595-0.56651.4808-1.19952.29470.0128-0.16290.14780.03270.00980.06480.0293-0.59010.01780.11620.0187-0.02460.2612-0.07130.1801-11.2554-16.275719.5991
27.17541.4282-2.11631.7422-1.23663.9151-0.10530.0786-0.0435-0.04150.0460.19980.1096-0.61130.06310.13190.0242-0.04110.269-0.05160.1905-14.8024-16.876814.0894
35.2794-1.1728-0.43940.94510.32081.1395-0.02240.20270.2029-0.0909-0.01360.0942-0.1029-0.13380.04690.1731-0.0039-0.05080.1061-0.01540.1631-0.441-13.182111.661
45.711-4.1691-2.25775.13252.83173.2941-0.08890.1780.1192-0.1835-0.05010.2089-0.2831-0.08040.10620.15060.0106-0.05780.0985-0.00230.1794.7525-10.363911.3227
52.4107-0.2872-0.40050.99140.50821.8481-0.0623-0.13150.22590.0332-0.09950.2221-0.1956-0.20150.08830.14770.0298-0.03220.1185-0.05960.20315.1768-11.335223.2624
62.4679-0.3082-0.00473.9972-0.59515.2277-0.2084-0.0268-0.8903-0.3612-0.1030.09141.1362-0.28130.09110.3629-0.04120.05230.19830.01670.41173.1045-33.947524.7233
70.69730.54850.51940.6745-0.05311.2215-0.026-0.38620.1080.01450.00120.1594-0.0609-0.30560.03490.14790.00440.0210.2155-0.05310.15682.453-17.728332.1768
83.5468-0.5728-0.77013.96360.54653.6621-0.07850.29480.0982-0.22170.0823-0.2259-0.10920.5063-0.01930.1169-0.07710.0020.21110.02010.134441.7311-12.38398.5713
95.07961.23391.71142.55530.24411.8303-0.04630.30680.3298-0.0817-0.1078-0.4165-0.36470.40460.13380.2268-0.0842-0.01170.36010.09280.193941.1845-8.01522.8803
103.6614-1.1129-0.47711.36650.31991.1221-0.06050.33830.2141-0.07970.0288-0.0045-0.18590.0576-0.00090.1495-0.0318-0.03110.11170.01440.120624.3471-12.78166.6017
113.3083-0.55960.22671.60680.31451.7959-0.0397-0.07410.01740.045-0.0081-0.0375-0.03270.09070.04490.1059-0.0317-0.01450.08780.01260.100426.5254-16.565518.029
121.13230.4325-0.25472.18140.50211.27340.0149-0.09010.3234-0.0147-0.0150.0604-0.33050.13950.00050.2078-0.0422-0.04530.145-0.0190.200831.2078-3.214325.223
133.0644-0.5139-2.19452.4967-2.87826.6475-0.1022-0.42770.04710.33240.08710.26810.2076-0.6569-0.02090.24130.03830.04820.5192-0.06670.1816-5.8941-19.580356.1145
140.8035-0.349-0.9890.5560.47121.235-0.0529-0.69330.30760.1729-0.11130.1611-0.4511-0.62520.07480.35170.0560.03450.5688-0.14430.2572-4.1642-12.079657.8739
155.15161.40920.06311.00960.26221.4040.0359-0.58280.00990.1805-0.07830.0187-0.0894-0.07450.06670.2458-0.00460.0130.3566-0.04960.12389.7547-16.571657.0295
161.7165-0.24860.2630.64050.04912.23510.0252-0.39920.09380.1192-0.10220.0602-0.1337-0.04990.06750.17420.00280.00740.2636-0.02330.121111.1272-16.737348.588
171.74080.1998-0.46762.403-0.75091.08520.0254-0.17590.36290.2545-0.03190.0524-0.2773-0.10190.01360.23440.0435-0.03440.2491-0.08360.21825.5208-4.894938.6302
183.8375-0.06090.77933.3920.13544.33020.114-0.5172-0.2170.0722-0.2346-0.30560.02730.69380.12750.15480.0477-0.03170.57110.10280.213147.8012-20.480241.6141
192.0313-2.6911-1.10113.97131.16854.8358-0.0053-0.541-1.19850.5149-0.0833-0.35880.46211.26460.08040.38060.0504-0.0580.6320.10890.70352.0093-27.784347.0896
202.86280.4986-0.50520.9248-1.09781.29820.0124-0.66630.11760.262-0.1552-0.197-0.3380.56350.11140.2613-0.0476-0.05910.60370.00630.222945.8532-15.265451.2625
214.80851.5637-1.45940.6483-0.1021.48470.0567-0.354-0.79280.10270.0055-0.04310.28830.24850.00630.30.02840.0030.36740.17190.325522.7729-31.256950.6248
221.6233-0.0945-0.12780.4608-0.1131.63110.0309-0.3834-0.02920.1156-0.0828-0.0662-0.10140.24580.05580.1728-0.0088-0.02350.26740.0140.128231.5406-16.451444.2099
234.6155-0.95710.73373.41-2.8373.37670.1519-0.1177-0.75650.0403-0.18760.02121.51080.103-0.05370.41810.07970.08370.22830.0460.347129.3664-38.738136.7976
241.61-0.6448-0.00210.8354-0.10431.6875-0.0779-0.1033-0.2572-0.06030.0189-0.07020.17160.29910.05040.16230.0411-0.00030.18240.03590.141933.0914-23.054229.609
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection detailsAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1chain 'A' and (resid 3 through 40 )A3 - 40
2X-RAY DIFFRACTION2chain 'A' and (resid 41 through 63 )A41 - 63
3X-RAY DIFFRACTION3chain 'A' and (resid 64 through 102 )A64 - 102
4X-RAY DIFFRACTION4chain 'A' and (resid 103 through 130 )A103 - 130
5X-RAY DIFFRACTION5chain 'A' and (resid 131 through 188 )A131 - 188
6X-RAY DIFFRACTION6chain 'A' and (resid 189 through 219 )A189 - 219
7X-RAY DIFFRACTION7chain 'A' and (resid 220 through 254 )A220 - 254
8X-RAY DIFFRACTION8chain 'B' and (resid 0 through 40 )B0 - 40
9X-RAY DIFFRACTION9chain 'B' and (resid 41 through 63 )B41 - 63
10X-RAY DIFFRACTION10chain 'B' and (resid 64 through 130 )B64 - 130
11X-RAY DIFFRACTION11chain 'B' and (resid 131 through 188 )B131 - 188
12X-RAY DIFFRACTION12chain 'B' and (resid 189 through 254 )B189 - 254
13X-RAY DIFFRACTION13chain 'C' and (resid -4 through 15 )C-4 - 15
14X-RAY DIFFRACTION14chain 'C' and (resid 16 through 63 )C16 - 63
15X-RAY DIFFRACTION15chain 'C' and (resid 64 through 102 )C64 - 102
16X-RAY DIFFRACTION16chain 'C' and (resid 103 through 199 )C103 - 199
17X-RAY DIFFRACTION17chain 'C' and (resid 200 through 254 )C200 - 254
18X-RAY DIFFRACTION18chain 'D' and (resid 2 through 36 )D2 - 36
19X-RAY DIFFRACTION19chain 'D' and (resid 37 through 50 )D37 - 50
20X-RAY DIFFRACTION20chain 'D' and (resid 51 through 86 )D51 - 86
21X-RAY DIFFRACTION21chain 'D' and (resid 87 through 102 )D87 - 102
22X-RAY DIFFRACTION22chain 'D' and (resid 103 through 193 )D103 - 193
23X-RAY DIFFRACTION23chain 'D' and (resid 194 through 209 )D194 - 209
24X-RAY DIFFRACTION24chain 'D' and (resid 210 through 254 )D210 - 254

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