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- PDB-1vl6: Crystal structure of NAD-dependent malic enzyme (TM0542) from The... -

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Basic information

Entry
Database: PDB / ID: 1vl6
TitleCrystal structure of NAD-dependent malic enzyme (TM0542) from Thermotoga maritima at 2.61 A resolution
Componentsmalate oxidoreductase
KeywordsOXIDOREDUCTASE / TM0542 / NAD-DEPENDENT MALIC ENZYME / STRUCTURAL GENOMICS / JCSG / PROTEIN STRUCTURE INITIATIVE / PSI / Joint Center for Structural Genomics
Function / homology
Function and homology information


malic enzyme activity / oxidoreductase activity, acting on the CH-OH group of donors, NAD or NADP as acceptor / NAD binding / metal ion binding
Similarity search - Function
Malic enzyme, NAD-binding domain, bacterial type / Malic enzyme, N-terminal domain / Malic oxidoreductase / Malic enzyme, N-terminal domain / Malic enzyme, N-terminal domain / Malic enzyme, NAD-binding / Malic enzyme, N-terminal domain superfamily / Malic enzyme, N-terminal domain / Malic enzyme, NAD binding domain / Malic enzyme, NAD binding domain ...Malic enzyme, NAD-binding domain, bacterial type / Malic enzyme, N-terminal domain / Malic oxidoreductase / Malic enzyme, N-terminal domain / Malic enzyme, N-terminal domain / Malic enzyme, NAD-binding / Malic enzyme, N-terminal domain superfamily / Malic enzyme, N-terminal domain / Malic enzyme, NAD binding domain / Malic enzyme, NAD binding domain / Aminoacid dehydrogenase-like, N-terminal domain superfamily / NAD(P)-binding Rossmann-like Domain / NAD(P)-binding domain superfamily / Rossmann fold / 3-Layer(aba) Sandwich / Alpha Beta
Similarity search - Domain/homology
Malate oxidoreductase
Similarity search - Component
Biological speciesThermotoga maritima (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MAD / Resolution: 2.61 Å
AuthorsJoint Center for Structural Genomics (JCSG)
CitationJournal: To be published
Title: Crystal structure of NAD-dependent malic enzyme (TM0542) from Thermotoga maritima at 2.61 A resolution
Authors: Joint Center for Structural Genomics (JCSG)
History
DepositionJul 13, 2004Deposition site: RCSB / Processing site: RCSB
Revision 1.0Aug 17, 2004Provider: repository / Type: Initial release
Revision 1.1Apr 26, 2008Group: Version format compliance
Revision 1.2Jul 13, 2011Group: Advisory / Source and taxonomy / Version format compliance
Revision 1.3Jan 25, 2023Group: Database references / Derived calculations / Category: database_2 / struct_conn / struct_ref_seq_dif
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession ..._database_2.pdbx_DOI / _database_2.pdbx_database_accession / _struct_conn.pdbx_leaving_atom_flag / _struct_ref_seq_dif.details
Remark 999 SEQUENCE FOR THE VAL/MET CONFLICT AT RESIDUE 1: THIS GENE PRODUCT WAS EXPRESSED AS A FUSION ... SEQUENCE FOR THE VAL/MET CONFLICT AT RESIDUE 1: THIS GENE PRODUCT WAS EXPRESSED AS A FUSION PROTEIN, SO THE ORIGINAL INITIATION CODON WAS TRANSLATED AS VAL INSTEAD OF MET. THERE IS A POINT MUTATION, ILE TO VAL, AT POSITION 6. IT WAS AN ERROR INTRODUCED DURING CLONING. THIS IS SUPPORTED BY THE ELECTRON DENSITY WHICH CLEARLY SHOW VAL RESIDUES AT THE POSITION 6 OF ALL FOUR CHAINS.

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: malate oxidoreductase
B: malate oxidoreductase
C: malate oxidoreductase
D: malate oxidoreductase


Theoretical massNumber of molelcules
Total (without water)171,3404
Polymers171,3404
Non-polymers00
Water79344
1


  • Idetical with deposited unit
  • defined by author&software
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area20200 Å2
ΔGint-172 kcal/mol
Surface area51230 Å2
MethodPISA
Unit cell
Length a, b, c (Å)143.963, 143.963, 163.428
Angle α, β, γ (deg.)90.00, 90.00, 120.00
Int Tables number170
Space group name H-MP65
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11A
21B
31C
41D
12A
22B
32C
42D
13A
23C
33D
14A
24B
34C
15A
25C
35D
16A
26B
36C
46D

NCS domain segments:

Component-ID: 1 / Refine code: 3

Dom-IDEns-IDBeg label comp-IDEnd label comp-IDAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
11ASPGLNAA2 - 15614 - 168
21ASPGLNBB2 - 15614 - 168
31ASPGLNCC2 - 15614 - 168
41ASPGLNDD2 - 15614 - 168
12GLNGLUAA157 - 175169 - 187
22GLNGLUBB157 - 175169 - 187
32GLNGLUCC157 - 175169 - 187
42GLNGLUDD157 - 175169 - 187
13LYSVALAA176 - 185188 - 197
23LYSVALCC176 - 185188 - 197
33LYSVALDD176 - 185188 - 197
14ASNILEAA186 - 235198 - 247
24ASNILEBB186 - 235198 - 247
34ASNILECC186 - 235198 - 247
15THRASNAA236 - 310248 - 322
25THRASNCC236 - 310248 - 322
35THRASNDD236 - 310248 - 322
16GLNSERAA311 - 375323 - 387
26GLNSERBB311 - 375323 - 387
36GLNSERCC311 - 375323 - 387
46GLNSERDD311 - 375323 - 387

NCS ensembles :
ID
1
2
3
4
5
6

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Components

#1: Protein
malate oxidoreductase


Mass: 42835.117 Da / Num. of mol.: 4
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Thermotoga maritima (bacteria) / Strain: MSB8 / Gene: TM0542 / Production host: Escherichia coli (E. coli) / References: UniProt: Q9WZ12
#2: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 44 / 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: 2

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Sample preparation

Crystal
IDDensity Matthews3/Da)Density % sol (%)
12.8556.87
23.1260.31
Crystal grow
Temperature (K)Crystal-IDMethodDetails
2771vapor diffusion, sitting drop, nanodrop0.025M Citric Acid, 0.075M Citrate_Na3, 6.00% NP_Peg 6000 , VAPOR DIFFUSION,SITTING DROP,NANODROP, temperature 277K
2772vapor diffusion, sitting drop, nanodrop0.025M Citric Acid, 0.075M Citrate_Na3, 6.00% NP_Peg 6000 , VAPOR DIFFUSION,SITTING DROP,NANODROP, temperature 277K

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Data collection

Diffraction
IDMean temperature (K)Crystal-ID
11001
21001
1,21
Diffraction source
SourceSiteBeamlineIDWavelengthWavelength (Å)
SYNCHROTRONALS 8.3.111.11587
SYNCHROTRONALS 8.3.121.019943, 0.979762, 0.979619
DetectorType: ADSC / Detector: CCD / Date: Jul 25, 2003
Radiation
IDMonochromatorProtocolMonochromatic (M) / Laue (L)Scattering typeWavelength-ID
1Double Crystal Si(111)SINGLE WAVELENGTHMx-ray1
2MADMx-ray2
Radiation wavelength
IDWavelength (Å)Relative weight
11.115871
21.0199431
30.9797621
40.9796191
ReflectionResolution: 2.6→47.12 Å / Num. obs: 57871 / % possible obs: 98.84 % / Redundancy: 3.26 % / Biso Wilson estimate: 74.19 Å2 / Rmerge(I) obs: 0.054 / Net I/σ(I): 16.77
Reflection shellResolution: 2.6→2.69 Å / Redundancy: 2.86 % / Rmerge(I) obs: 0.551 / Mean I/σ(I) obs: 1.84 / Num. unique all: 5591 / % possible all: 96.26

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Processing

Software
NameVersionClassification
DENZOdata reduction
MOSFLMdata reduction
SCALEPACKdata scaling
SOLVEphasing
SHARPphasing
REFMAC5.2.0003refinement
RefinementMethod to determine structure: MAD / Resolution: 2.61→47.12 Å / Cor.coef. Fo:Fc: 0.962 / Cor.coef. Fo:Fc free: 0.941 / SU B: 23.972 / SU ML: 0.225 / TLS residual ADP flag: LIKELY RESIDUAL / Cross valid method: THROUGHOUT / ESU R: 0.524 / ESU R Free: 0.286
Stereochemistry target values: MAXIMUM LIKELIHOOD WITH PHASES
Details: 1. HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS. 2. THE ELECTRON DENSITY IN THE REGIONS OF B243-B254, B267-B268 AND B287-B306 ARE POOR, BUT THE DENSITY STILL SHOW CLEARLY THE MAIN CHAIN ...Details: 1. HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS. 2. THE ELECTRON DENSITY IN THE REGIONS OF B243-B254, B267-B268 AND B287-B306 ARE POOR, BUT THE DENSITY STILL SHOW CLEARLY THE MAIN CHAIN TRACES, SO THEY WERE STILL INCLUDED IN THIS MODEL.
RfactorNum. reflection% reflectionSelection details
Rfree0.23929 2333 4.1 %RANDOM
Rwork0.19189 ---
obs0.1938 55267 98.53 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.2 Å / Solvent model: BABINET MODEL WITH MASK
Displacement parametersBiso mean: 61.902 Å2
Baniso -1Baniso -2Baniso -3
1-1.1 Å20.55 Å20 Å2
2--1.1 Å20 Å2
3----1.64 Å2
Refinement stepCycle: LAST / Resolution: 2.61→47.12 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms11096 0 0 44 11140
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0170.02211343
X-RAY DIFFRACTIONr_bond_other_d0.0020.0210623
X-RAY DIFFRACTIONr_angle_refined_deg1.6581.96115422
X-RAY DIFFRACTIONr_angle_other_deg0.933324642
X-RAY DIFFRACTIONr_dihedral_angle_1_deg6.6551486
X-RAY DIFFRACTIONr_dihedral_angle_2_deg38.55625.045448
X-RAY DIFFRACTIONr_dihedral_angle_3_deg17.945151862
X-RAY DIFFRACTIONr_dihedral_angle_4_deg17.6681550
X-RAY DIFFRACTIONr_chiral_restr0.0960.21810
X-RAY DIFFRACTIONr_gen_planes_refined0.0060.0212724
X-RAY DIFFRACTIONr_gen_planes_other0.0020.022119
X-RAY DIFFRACTIONr_nbd_refined0.2360.22569
X-RAY DIFFRACTIONr_nbd_other0.1790.211098
X-RAY DIFFRACTIONr_nbtor_other0.0910.27174
X-RAY DIFFRACTIONr_xyhbond_nbd_refined0.150.2248
X-RAY DIFFRACTIONr_symmetry_vdw_refined0.40.217
X-RAY DIFFRACTIONr_symmetry_vdw_other0.2740.236
X-RAY DIFFRACTIONr_symmetry_hbond_refined0.0940.27
X-RAY DIFFRACTIONr_mcbond_it1.52137552
X-RAY DIFFRACTIONr_mcbond_other0.49233009
X-RAY DIFFRACTIONr_mcangle_it2.64511927
X-RAY DIFFRACTIONr_scbond_it5.884167
X-RAY DIFFRACTIONr_scangle_it8.133113495
X-RAY DIFFRACTIONr_nbtor_refined0.1890.25819
X-RAY DIFFRACTIONr_xyhbond_nbd_other0.0650.21
Refine LS restraints NCS

Refine-ID: X-RAY DIFFRACTION

Ens-IDDom-IDAuth asym-IDNumberTypeRms dev position (Å)Weight position
11A904tight positional0.050.05
12B904tight positional0.050.05
13C904tight positional0.040.05
14D904tight positional0.050.05
11A1333loose positional0.445
12B1333loose positional0.475
13C1333loose positional0.55
14D1333loose positional0.495
11A904tight thermal0.150.5
12B904tight thermal0.140.5
13C904tight thermal0.120.5
14D904tight thermal0.140.5
11A1333loose thermal2.5610
12B1333loose thermal2.4810
13C1333loose thermal2.5110
14D1333loose thermal2.4310
21A113tight positional0.040.05
22B113tight positional0.040.05
23C113tight positional0.050.05
24D113tight positional0.040.05
21A130loose positional0.225
22B130loose positional0.315
23C130loose positional0.45
24D130loose positional0.235
21A113tight thermal0.180.5
22B113tight thermal0.110.5
23C113tight thermal0.10.5
24D113tight thermal0.140.5
21A130loose thermal2.1510
22B130loose thermal2.0410
23C130loose thermal2.2610
24D130loose thermal1.4810
31A60tight positional0.050.05
32C60tight positional0.050.05
33D60tight positional0.040.05
31A87loose positional0.345
32C87loose positional0.455
33D87loose positional0.315
31A60tight thermal0.160.5
32C60tight thermal0.130.5
33D60tight thermal0.150.5
31A87loose thermal2.7710
32C87loose thermal2.9710
33D87loose thermal3.8510
41A294tight positional0.060.05
42B294tight positional0.050.05
43C294tight positional0.060.05
41A415loose positional0.525
42B415loose positional0.415
43C415loose positional0.545
41A294tight thermal0.160.5
42B294tight thermal0.120.5
43C294tight thermal0.130.5
41A415loose thermal3.4110
42B415loose thermal2.9510
43C415loose thermal2.6910
51A437tight positional0.050.05
52C437tight positional0.050.05
53D437tight positional0.040.05
51A609loose positional0.555
52C609loose positional0.795
53D609loose positional0.565
51A437tight thermal0.150.5
52C437tight thermal0.120.5
53D437tight thermal0.140.5
51A609loose thermal3.0410
52C609loose thermal3.0510
53D609loose thermal2.5110
61A377tight positional0.060.05
62B377tight positional0.080.05
63C377tight positional0.070.05
64D377tight positional0.060.05
61A540loose positional0.555
62B540loose positional0.745
63C540loose positional0.575
64D540loose positional0.535
61A377tight thermal0.160.5
62B377tight thermal0.120.5
63C377tight thermal0.110.5
64D377tight thermal0.150.5
61A540loose thermal3.6710
62B540loose thermal3.7410
63C540loose thermal3.3110
64D540loose thermal3.1110
LS refinement shellResolution: 2.61→2.672 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.394 179 4.48 %
Rwork0.326 3819 -
obs--93.81 %
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)
11.8404-0.10310.05681.8361-0.26212.5428-0.10210.28480.5246-0.00130.14560.1757-0.0907-0.0432-0.0435-0.30540.04820.0117-0.24170.0702-0.1234-16.046849.145519.0726
22.7455-0.44471.2852.3602-0.392.16140.0483-0.3253-0.05520.29250.00770.19670.0961-0.2565-0.056-0.21980.010.04-0.25660.0117-0.4146-7.74522.192237.0643
31.8928-0.3048-0.64732.9760.5921.5440.01240.2580.1015-0.2701-0.02920.0921-0.08840.04770.0168-0.26340.0160.0289-0.24660.0373-0.4021-8.933229.367116.406
42.3312-0.3718-0.13591.9290.01911.4061-0.03980.27350.5432-0.20190.0970.3329-0.1441-0.0744-0.0572-0.26950.03260.0037-0.25040.13520.1039-17.686158.438317.9036
510.622.2405-4.70693.5466-1.76655.29720.50792.0190.898-0.2605-0.00770.4465-1.0883-1.4861-0.50020.32790.38660.02530.32620.09270.2617-35.938282.99419.2463
67.7671-1.6566-0.90665.5914-1.31143.53670.16330.64470.5253-0.5729-0.2014-0.7736-0.3660.04670.03820.07160.06870.1329-0.16070.04210.3462-15.856378.316811.4574
73.19370.7021-0.48311.97450.21891.39470.09160.420.73070.03230.18770.1356-0.1102-0.2517-0.2793-0.1793-0.08290.0521-0.08590.29250.032110.203160.4584-4.0488
83.68031.0731-1.76712.7922-0.65584.29410.6492-1.1380.71860.9507-0.00180.5991-1.03250.6625-0.64740.3969-0.28750.31540.1589-0.00940.240825.525188.88674.1883
93.40160.5015-1.82664.07051.46013.25050.3905-0.23120.61150.24870.1240.9519-0.7383-0.2269-0.51450.10480.02660.32340.04870.35840.64935.609980.13871.6806
102.48690.684-0.37441.74780.33551.8098-0.04360.49940.4559-0.15540.21430.0723-0.0246-0.338-0.1707-0.2237-0.11390.0108-0.01930.245-0.15979.627150.7747-5.6852
113.21510.41281.8462.13850.87563.10620.03610.943-0.1565-0.35930.1644-0.21560.08391.1352-0.2005-0.0586-0.05340.0610.1769-0.0162-0.35394.666322.823-23.8468
122.15240.78191.57883.8751-0.511.6892-0.0280.07580.14290.1393-0.0374-0.0183-0.13540.13680.0655-0.2022-0.09390.0418-0.11680.0334-0.39774.909530.2439-3.3242
Refinement TLS group

Refine-ID: X-RAY DIFFRACTION / Selection: ALL

IDRefine TLS-IDAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
11AA0 - 15612 - 168
22AA157 - 309169 - 321
33AA310 - 376322 - 388
44BB0 - 15612 - 168
55BB157 - 309169 - 321
66BB310 - 376322 - 388
77CC0 - 15612 - 168
88CC157 - 309169 - 321
99CC310 - 376322 - 388
1010DD0 - 15612 - 168
1111DD157 - 309169 - 321
1212DD310 - 376322 - 388

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