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- PDB-6uca: Crystal structure of human ZCCHC4 in complex with SAH -

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Basic information

Entry
Database: PDB / ID: 6uca
TitleCrystal structure of human ZCCHC4 in complex with SAH
ComponentsrRNA N6-adenosine-methyltransferase ZCCHC4
KeywordsTRANSFERASE / 28s RNA m6A methyltransferase
Function / homology
Function and homology information


rRNA (adenine-N6-)-methyltransferase activity / S-adenosyl-L-methionine binding / rRNA methylation / Transferases; Transferring one-carbon groups; Methyltransferases / positive regulation of translation / nucleic acid binding / nucleolus / zinc ion binding / cytoplasm
Similarity search - Function
Zinc finger CCHC domain-containing protein 4 / Methyltransferase EEF1AKMT1/ZCCHC4 / Probable N6-adenine methyltransferase / Zinc finger, GRF-type / GRF zinc finger / DHHC domain profile. / N-6 Adenine-specific DNA methylases signature. / DNA methylase, N-6 adenine-specific, conserved site
Similarity search - Domain/homology
S-ADENOSYL-L-HOMOCYSTEINE / rRNA N6-adenosine-methyltransferase ZCCHC4
Similarity search - Component
Biological speciesHomo sapiens (human)
MethodX-RAY DIFFRACTION / SYNCHROTRON / SAD / Resolution: 3.103 Å
AuthorsLu, J.W. / Ren, W.D. / Huang, M.J. / Gao, L. / Li, D.X. / Wang, G.G. / Song, J.
Funding support United States, 1items
OrganizationGrant numberCountry
National Institutes of Health/National Human Genome Research Institute (NIH/NHGRI) United States
CitationJournal: Nat Commun / Year: 2019
Title: Structure and regulation of ZCCHC4 in m6A-methylation of 28S rRNA.
Authors: Ren, W. / Lu, J. / Huang, M. / Gao, L. / Li, D. / Wang, G.G. / Song, J.
History
DepositionSep 15, 2019Deposition site: RCSB / Processing site: RCSB
Revision 1.0Oct 16, 2019Provider: repository / Type: Initial release
Revision 1.1Dec 18, 2019Group: Author supporting evidence / Category: pdbx_audit_support / Item: _pdbx_audit_support.funding_organization
Revision 1.2Apr 29, 2020Group: Database references / Category: citation / citation_author
Item: _citation.country / _citation.journal_abbrev ..._citation.country / _citation.journal_abbrev / _citation.journal_id_CSD / _citation.journal_id_ISSN / _citation.journal_volume / _citation.page_first / _citation.page_last / _citation.pdbx_database_id_DOI / _citation.pdbx_database_id_PubMed / _citation.title / _citation.year / _citation_author.identifier_ORCID / _citation_author.name
Revision 1.3Mar 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

Downloads & links

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Assembly

Deposited unit
A: rRNA N6-adenosine-methyltransferase ZCCHC4
B: rRNA N6-adenosine-methyltransferase ZCCHC4
C: rRNA N6-adenosine-methyltransferase ZCCHC4
D: rRNA N6-adenosine-methyltransferase ZCCHC4
E: rRNA N6-adenosine-methyltransferase ZCCHC4
F: rRNA N6-adenosine-methyltransferase ZCCHC4
hetero molecules


Theoretical massNumber of molelcules
Total (without water)309,54748
Polymers304,8866
Non-polymers4,66142
Water0
1
A: rRNA N6-adenosine-methyltransferase ZCCHC4
hetero molecules


Theoretical massNumber of molelcules
Total (without water)51,5918
Polymers50,8141
Non-polymers7777
Water0
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
2
B: rRNA N6-adenosine-methyltransferase ZCCHC4
hetero molecules


Theoretical massNumber of molelcules
Total (without water)51,5918
Polymers50,8141
Non-polymers7777
Water0
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
3
C: rRNA N6-adenosine-methyltransferase ZCCHC4
hetero molecules


Theoretical massNumber of molelcules
Total (without water)51,5918
Polymers50,8141
Non-polymers7777
Water0
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
4
D: rRNA N6-adenosine-methyltransferase ZCCHC4
hetero molecules


Theoretical massNumber of molelcules
Total (without water)51,5918
Polymers50,8141
Non-polymers7777
Water0
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
5
E: rRNA N6-adenosine-methyltransferase ZCCHC4
hetero molecules


Theoretical massNumber of molelcules
Total (without water)51,5918
Polymers50,8141
Non-polymers7777
Water0
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
6
F: rRNA N6-adenosine-methyltransferase ZCCHC4
hetero molecules


Theoretical massNumber of molelcules
Total (without water)51,5918
Polymers50,8141
Non-polymers7777
Water0
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Unit cell
Length a, b, c (Å)136.567, 290.380, 83.529
Angle α, β, γ (deg.)90.000, 90.000, 90.000
Int Tables number18
Space group name H-MP21212

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Components

#1: Protein
rRNA N6-adenosine-methyltransferase ZCCHC4 / Zinc finger CCHC domain-containing protein 4


Mass: 50814.344 Da / Num. of mol.: 6
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Homo sapiens (human) / Gene: ZCCHC4 / Production host: Escherichia coli BL21(DE3) (bacteria)
References: UniProt: Q9H5U6, Transferases; Transferring one-carbon groups; Methyltransferases
#2: Chemical
ChemComp-SAH / S-ADENOSYL-L-HOMOCYSTEINE / S-Adenosyl-L-homocysteine


Type: L-peptide linking / Mass: 384.411 Da / Num. of mol.: 6 / Source method: obtained synthetically / Formula: C14H20N6O5S / Feature type: SUBJECT OF INVESTIGATION
#3: Chemical...
ChemComp-ZN / ZINC ION


Mass: 65.409 Da / Num. of mol.: 36 / Source method: obtained synthetically / Formula: Zn / Feature type: SUBJECT OF INVESTIGATION
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: 2.94 Å3/Da / Density % sol: 58.22 %
Crystal growTemperature: 285 K / Method: vapor diffusion, sitting drop / pH: 7.6
Details: 0.03 M citric acid, 0.07 M Bis-Tris propane, pH 7.6, 11-12% PEG3350, 10% glycerol, 5 mM TCEP, pH 7.0

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Data collection

DiffractionMean temperature: 100 K / Serial crystal experiment: N
Diffraction sourceSource: SYNCHROTRON / Site: ALS / Beamline: 5.0.2 / Wavelength: 1.283 Å
DetectorType: DECTRIS PILATUS 6M / Detector: PIXEL / Date: May 29, 2019
RadiationMonochromator: double crystal Si(111) / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 1.283 Å / Relative weight: 1
ReflectionResolution: 3.1→50 Å / Num. obs: 60780 / % possible obs: 99.8 % / Redundancy: 5.7 % / Rmerge(I) obs: 0.126 / Rpim(I) all: 0.057 / Rrim(I) all: 0.139 / Χ2: 1.049 / Net I/σ(I): 6
Reflection shell

Diffraction-ID: 1

Resolution (Å)Redundancy (%)Rmerge(I) obsNum. unique obsCC1/2Rpim(I) allRrim(I) allΧ2% possible all
3.1-3.215.50.75259870.9190.3490.8310.51299.9
3.21-3.345.80.56259960.9430.2520.6180.54899.8
3.34-3.495.90.4159880.9650.1820.450.63699.9
3.49-3.685.80.30960110.9710.1390.340.76199.8
3.68-3.915.50.19960170.9840.0930.2210.91799.7
3.91-4.215.80.15660590.990.0710.1721.04399.9
4.21-4.635.90.12260750.9920.0550.1341.32699.9
4.63-5.35.50.10560880.9920.0490.1161.44899.9
5.3-6.675.90.09861870.9920.0440.1081.416100
6.67-505.40.06664310.9960.030.0731.85399.7

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Processing

Software
NameVersionClassification
PHENIX1.15.2_3472refinement
SCALEPACKdata scaling
PDB_EXTRACT3.25data extraction
HKL-3000data reduction
CRANK2phasing
RefinementMethod to determine structure: SAD / Resolution: 3.103→47.683 Å / SU ML: 0.42 / Cross valid method: THROUGHOUT / σ(F): 1.35 / Phase error: 31.82 / Stereochemistry target values: ML
RfactorNum. reflection% reflection
Rfree0.2813 3795 3.3 %
Rwork0.249 111100 -
obs0.25 60686 98.96 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL
Displacement parametersBiso max: 390.44 Å2 / Biso mean: 105.472 Å2 / Biso min: 28.5 Å2
Refinement stepCycle: final / Resolution: 3.103→47.683 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms18478 0 156 0 18634
Biso mean--76.97 --
Num. residues----2445
LS refinement shell

Refine-ID: X-RAY DIFFRACTION / Rfactor Rfree error: 0

Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection Rwork% reflection obs (%)
3.103-3.14220.37861310.3682376491
3.1422-3.18350.3281390.3425413399
3.1835-3.22710.37551440.3194411499
3.2271-3.27320.26311360.2927409299
3.2732-3.32210.29491470.29544202100
3.3221-3.3740.32531360.2959407999
3.374-3.42930.36971390.28794131100
3.4293-3.48840.30971460.2923422499
3.4884-3.55180.33921390.30434041100
3.5518-3.62010.32221440.2817417099
3.6201-3.69390.28221420.2793409999
3.6939-3.77420.28811390.2537413099
3.7742-3.8620.28581400.2425409299
3.862-3.95850.27661400.24394146100
3.9585-4.06550.32551390.24664141100
4.0655-4.18510.26951390.2331415899
4.1851-4.32010.29311420.2221412299
4.3201-4.47440.23511400.21914143100
4.4744-4.65340.27131430.21234147100
4.6534-4.86490.25021410.21764151100
4.8649-5.12120.28921440.2221407599
5.1212-5.44160.26171310.23784138100
5.4416-5.8610.26111430.2454144100
5.861-6.44960.30031470.2731417699
6.4496-7.37990.3641330.2692409599
7.3799-9.28660.27911420.2288416299
9.2866-47.6830.19941490.2172403198
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)
11.6528-0.3813-0.03651.40580.02013.4776-0.40360.4689-0.8595-0.5667-0.09750.61820.9713-1.78990.51990.8936-0.40770.17241.414-0.30060.8122131.147571.549833.0607
22.64620.25720.21414.55560.89364.7795-0.1317-0.489-0.11160.54680.0433-0.4611-0.0057-0.39380.22160.30520.1291-0.09120.5840.04130.2935149.727682.840158.235
36.0501-1.7306-1.75955.1473-0.80959.17060.09120.19080.1531-0.208-0.1424-0.8398-0.29660.03020.07830.23740.00270.060.295-0.03540.4315156.903282.89540.32
43.8811-1.4227-0.71644.70842.06062.792-0.2940.1584-1.1089-0.4009-0.0467-0.95820.80850.06020.33440.7938-0.07690.41520.6088-0.13750.8717164.072263.643830.4996
55.0023-0.6077-2.01581.97432.21213.05140.1791.89741.0703-0.6257-0.32330.262-0.7141-0.44130.0821.41720.250.30051.58530.56981.307948.216227.6669-11.9431
61.0447-0.02490.37191.9141-0.32820.75190.6630.35631.6603-0.5422-0.0533-0.0002-0.35320.2656-0.28871.090.37040.9262-0.20850.34063.032254.457833.1547.3399
70.7416-0.89160.95051.2914-0.6052.4462-0.01111.03171.6172-0.89480.0588-0.4014-0.52030.44050.04311.52320.2671.01411.10250.98592.524669.679933.9876-7.36
87.3777-1.75821.61671.6509-1.62721.62041.22150.64821.2998-0.5536-0.4865-0.25510.35260.1596-0.65291.08340.1960.3840.61360.03790.923277.145612.37015.1787
99.2976-0.8418-0.39172.26930.9430.393-0.7342-1.5286-0.57870.89930.5956-0.04680.34990.36510.11763.3169-0.37980.16891.14180.06960.824590.062420.691559.0569
105.8828-1.3973-0.66136.14970.4323.5899-0.0674-0.13560.39440.832-0.4563-0.59620.7012-0.33920.38570.7086-0.21940.14720.4237-0.10030.645994.44538.495532.4449
113.53190.0138-1.53750.92320.39583.29570.1695-0.02630.01530.6631-0.32280.74780.3124-0.70410.08941.0151-0.5210.60960.9135-0.36410.815175.90934.392136.1908
121.12380.2076-0.68440.68170.58732.22680.179-0.1968-0.0240.8243-0.08560.46260.3077-0.3450.20191.8629-0.71150.86181.0038-0.18920.776473.0133.119350.2663
133.60441.1436-0.56971.8739-1.70531.61490.25010.166-0.5650.4272-0.25250.55150.6388-1.19920.05321.614-0.97270.96391.3198-0.51341.792358.74120.764537.4595
140.49250.08130.150.1384-0.06140.10081.4482-0.76650.6198-1.5070.252-0.51171.86730.7312-1.6492.62590.1554-0.25351.1644-0.62253.062558.9792-4.046742.2636
152.61890.04690.68982.0511-1.12154.4852-0.31130.3001-0.01440.0255-0.197-0.15580.56310.78060.46980.51130.06970.12910.54950.02980.427129.419478.5324-3.7116
165.27571.12490.28874.1604-0.37958.6735-0.2398-0.50250.24420.5664-0.01880.5279-0.3607-0.08830.18080.44170.0550.19140.4650.05730.3119.688484.74665.4352
172.29981.4379-0.39535.1341-3.1892.9798-0.28830.1379-0.8679-0.29130.09990.03940.5052-0.25380.12330.7851-0.05130.39540.67090.06260.8372107.778760.93229.9316
186.79385.2840.80515.63190.47645.22350.9489-0.77690.4880.4795-0.0538-0.6757-0.49650.5909-0.93970.9695-0.08470.28550.7183-0.16962.1157127.355127.19731.6175
195.03060.92510.53561.13151.50583.2210.8335-0.3213-0.29560.5633-0.4815-0.63660.3694-0.041-0.41261.1203-0.12350.10060.3861-0.04761.014899.423213.925726.0902
202.9345-0.64230.5920.6201-0.25195.098-0.04240.7043-0.5897-1.2625-0.1321-1.25890.9909-0.0799-0.02641.7472-0.08430.44180.5788-0.12051.2267107.256414.65664.8229
213.7542-0.38640.57360.0724-0.08134.35960.13630.4553-0.5483-1.094-0.0511-1.40110.95410.59130.03281.42580.1530.37140.5537-0.06891.9826119.00813.481913.1822
221.63450.96531.20180.8634-0.183.53-0.14230.49990.27-0.9931-0.1324-1.3061-0.16830.48020.30781.5414-0.02570.52030.5870.00511.7015115.92329.96375.005
231.6558-1.40060.27072.63871.80942.88460.25250.47510.74170.41490.1467-0.14030.5711-0.3245-0.56311.6186-0.23570.4780.68790.00121.2087111.732527.3839-1.743
241.11441.2217-0.47384.44721.10022.66390.58040.32391.0522-0.59670.4922-0.4282-0.5995-0.1714-0.99441.80130.08740.88630.87510.16731.8757118.008842.8397-6.6704
256.738-0.80640.45140.5028-1.12242.84821.22910.64450.7129-1.9634-0.49680.3783-0.6371-0.9107-0.71511.3830.5083-0.07050.94020.11892.32099.802130.23978.8924
265.3068-2.9049-0.83875.07670.8961.15930.72530.66370.7567-1.1011-0.71410.7906-0.1498-0.258-0.14820.85880.08020.02150.41020.13151.25433.910414.068912.1324
272.81611.17630.04290.86841.11433.39410.1108-0.35970.17930.8297-0.22681.46850.4124-0.29030.10530.9715-0.10970.45850.5258-0.00222.134818.179616.10828.9701
283.2679-2.6142-2.0532.56812.00891.6027-0.0339-0.84360.25940.94790.10480.94510.0410.2702-0.11191.2745-0.18360.64380.8885-0.13031.523918.65936.496745.1516
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection detailsAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1chain 'A' and (resid 28 through 97 )A28 - 97
2X-RAY DIFFRACTION2chain 'A' and (resid 98 through 176 )A98 - 176
3X-RAY DIFFRACTION3chain 'A' and (resid 177 through 283 )A177 - 283
4X-RAY DIFFRACTION4chain 'A' and (resid 284 through 440 )A284 - 440
5X-RAY DIFFRACTION5chain 'B' and (resid 28 through 116 )B28 - 116
6X-RAY DIFFRACTION6chain 'B' and (resid 117 through 252 )B117 - 252
7X-RAY DIFFRACTION7chain 'B' and (resid 253 through 311 )B253 - 311
8X-RAY DIFFRACTION8chain 'B' and (resid 312 through 438 )B312 - 438
9X-RAY DIFFRACTION9chain 'C' and (resid 36 through 98 )C36 - 98
10X-RAY DIFFRACTION10chain 'C' and (resid 99 through 159 )C99 - 159
11X-RAY DIFFRACTION11chain 'C' and (resid 160 through 241 )C160 - 241
12X-RAY DIFFRACTION12chain 'C' and (resid 242 through 297 )C242 - 297
13X-RAY DIFFRACTION13chain 'C' and (resid 298 through 415 )C298 - 415
14X-RAY DIFFRACTION14chain 'C' and (resid 416 through 442 )C416 - 442
15X-RAY DIFFRACTION15chain 'D' and (resid 25 through 200 )D25 - 200
16X-RAY DIFFRACTION16chain 'D' and (resid 201 through 311 )D201 - 311
17X-RAY DIFFRACTION17chain 'D' and (resid 312 through 440 )D312 - 440
18X-RAY DIFFRACTION18chain 'E' and (resid 28 through 97 )E28 - 97
19X-RAY DIFFRACTION19chain 'E' and (resid 98 through 176 )E98 - 176
20X-RAY DIFFRACTION20chain 'E' and (resid 177 through 213 )E177 - 213
21X-RAY DIFFRACTION21chain 'E' and (resid 214 through 311 )E214 - 311
22X-RAY DIFFRACTION22chain 'E' and (resid 312 through 347 )E312 - 347
23X-RAY DIFFRACTION23chain 'E' and (resid 348 through 372 )E348 - 372
24X-RAY DIFFRACTION24chain 'E' and (resid 373 through 439 )E373 - 439
25X-RAY DIFFRACTION25chain 'F' and (resid 28 through 84 )F28 - 84
26X-RAY DIFFRACTION26chain 'F' and (resid 85 through 176 )F85 - 176
27X-RAY DIFFRACTION27chain 'F' and (resid 177 through 344 )F177 - 344
28X-RAY DIFFRACTION28chain 'F' and (resid 345 through 439 )F345 - 439

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