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- PDB-4rmo: Crystal Structure of the CptIN Type III Toxin-Antitoxin System fr... -

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Basic information

Entry
Database: PDB / ID: 4rmo
TitleCrystal Structure of the CptIN Type III Toxin-Antitoxin System from Eubacterium rectale
Components
  • CptN Toxin
  • RNA (45-MER)
KeywordsToxin/RNA / protein-RNA complex / pseudoknot / RNA twist / toxin-antitoxin / type III / bacteriophage resistance / Toxin-RNA complex
Function / homologyThiol Ester Dehydrase; Chain A - #130 / Thiol Ester Dehydrase; Chain A / Roll / Alpha Beta / RNA / RNA (> 10) / Uncharacterized protein
Function and homology information
Biological speciesEubacterium rectale DSM 17629 (bacteria)
synthetic construct (others)
MethodX-RAY DIFFRACTION / SYNCHROTRON / SAD / Resolution: 2.2 Å
AuthorsRao, F. / Voss, J.E. / Short, F.L. / Luisi, B.F.
CitationJournal: Nucleic Acids Res. / Year: 2015
Title: Co-evolution of quaternary organization and novel RNA tertiary interactions revealed in the crystal structure of a bacterial protein-RNA toxin-antitoxin system.
Authors: Rao, F. / Short, F.L. / Voss, J.E. / Blower, T.R. / Orme, A.L. / Whittaker, T.E. / Luisi, B.F. / Salmond, G.P.
History
DepositionOct 21, 2014Deposition site: RCSB / Processing site: RCSB
Revision 1.0Sep 30, 2015Provider: repository / Type: Initial release
Revision 1.1Oct 14, 2015Group: Database references
Revision 1.2Nov 18, 2015Group: Database references

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: CptN Toxin
B: RNA (45-MER)
C: CptN Toxin
D: RNA (45-MER)
E: CptN Toxin
F: RNA (45-MER)
G: CptN Toxin
H: RNA (45-MER)
I: CptN Toxin
J: RNA (45-MER)
K: CptN Toxin
L: RNA (45-MER)
M: CptN Toxin
N: RNA (45-MER)
O: CptN Toxin
P: RNA (45-MER)
hetero molecules


Theoretical massNumber of molelcules
Total (without water)267,95164
Polymers266,02716
Non-polymers1,92448
Water23,8161322
1
A: CptN Toxin
B: RNA (45-MER)
E: CptN Toxin
F: RNA (45-MER)
hetero molecules


Theoretical massNumber of molelcules
Total (without water)66,94815
Polymers66,5074
Non-polymers44111
Water724
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area10810 Å2
ΔGint-143 kcal/mol
Surface area24740 Å2
MethodPISA
2
C: CptN Toxin
D: RNA (45-MER)
G: CptN Toxin
H: RNA (45-MER)
hetero molecules


Theoretical massNumber of molelcules
Total (without water)67,06818
Polymers66,5074
Non-polymers56114
Water724
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area10820 Å2
ΔGint-164 kcal/mol
Surface area25110 Å2
MethodPISA
3
I: CptN Toxin
J: RNA (45-MER)
K: CptN Toxin
L: RNA (45-MER)
hetero molecules


Theoretical massNumber of molelcules
Total (without water)67,02817
Polymers66,5074
Non-polymers52113
Water724
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area10750 Å2
ΔGint-149 kcal/mol
Surface area24740 Å2
MethodPISA
4
M: CptN Toxin
N: RNA (45-MER)
O: CptN Toxin
P: RNA (45-MER)
hetero molecules


Theoretical massNumber of molelcules
Total (without water)66,90714
Polymers66,5074
Non-polymers40110
Water724
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area10620 Å2
ΔGint-137 kcal/mol
Surface area25070 Å2
MethodPISA
Unit cell
Length a, b, c (Å)63.110, 185.891, 138.780
Angle α, β, γ (deg.)90.00, 92.63, 90.00
Int Tables number4
Space group name H-MP1211
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11A
21C
12A
22E
13A
23G
14A
24I
15A
25K
16A
26M
17A
27O
18B
28D
19B
29F
110B
210H
111B
211J
112B
212L
113B
213N
114B
214P
115C
215E
116C
216G
117C
217I
118C
218K
119C
219M
120C
220O
121D
221F
122D
222H
123D
223J
124D
224L
125D
225N
126D
226P
127E
227G
128E
228I
129E
229K
130E
230M
131E
231O
132F
232H
133F
233J
134F
234L
135F
235N
136F
236P
137G
237I
138G
238K
139G
239M
140G
240O
141H
241J
142H
242L
143H
243N
144H
244P
145I
245K
146I
246M
147I
247O
148J
248L
149J
249N
150J
250P
151K
251M
152K
252O
153L
253N
154L
254P
155M
255O
156N
256P

NCS domain segments:
Dom-IDComponent-IDEns-IDRefine codeAuth asym-IDAuth seq-ID
1010A1 - 155
2010C1 - 155
1020A1 - 155
2020E1 - 155
1030A1 - 155
2030G1 - 155
1040A1 - 155
2040I1 - 155
1050A1 - 155
2050K1 - 155
1060A1 - 155
2060M1 - 155
1070A1 - 155
2070O1 - 155
1080B1 - 45
2080D1 - 45
1090B1 - 45
2090F1 - 45
10100B1 - 45
20100H1 - 45
10110B1 - 45
20110J1 - 45
10120B1 - 45
20120L1 - 45
10130B1 - 45
20130N1 - 45
10140B1 - 45
20140P1 - 45
10150C1 - 155
20150E1 - 155
10160C1 - 155
20160G1 - 155
10170C1 - 155
20170I1 - 155
10180C1 - 155
20180K1 - 155
10190C1 - 155
20190M1 - 155
10200C1 - 155
20200O1 - 155
10210D1 - 45
20210F1 - 45
10220D1 - 45
20220H1 - 45
10230D1 - 45
20230J1 - 45
10240D1 - 45
20240L1 - 45
10250D1 - 45
20250N1 - 45
10260D1 - 45
20260P1 - 45
10270E1 - 155
20270G1 - 155
10280E1 - 155
20280I1 - 155
10290E1 - 155
20290K1 - 155
10300E1 - 155
20300M1 - 155
10310E1 - 155
20310O1 - 155
10320F1 - 45
20320H1 - 45
10330F1 - 45
20330J1 - 45
10340F1 - 45
20340L1 - 45
10350F1 - 45
20350N1 - 45
10360F1 - 45
20360P1 - 45
10370G1 - 155
20370I1 - 155
10380G1 - 155
20380K1 - 155
10390G1 - 155
20390M1 - 155
10400G1 - 155
20400O1 - 155
10410H1 - 45
20410J1 - 45
10420H1 - 45
20420L1 - 45
10430H1 - 45
20430N1 - 45
10440H1 - 45
20440P1 - 45
10450I1 - 155
20450K1 - 155
10460I1 - 155
20460M1 - 155
10470I1 - 155
20470O1 - 155
10480J1 - 45
20480L1 - 45
10490J1 - 45
20490N1 - 45
10500J1 - 45
20500P1 - 45
10510K1 - 155
20510M1 - 155
10520K1 - 155
20520O1 - 155
10530L1 - 45
20530N1 - 45
10540L1 - 45
20540P1 - 45
10550M1 - 155
20550O1 - 155
10560N1 - 45
20560P1 - 45

NCS ensembles :
ID
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56

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Components

#1: Protein
CptN Toxin


Mass: 18696.795 Da / Num. of mol.: 8
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Eubacterium rectale DSM 17629 (bacteria)
Strain: DSM 17629 / Gene: cptN, EUR_02830 / Plasmid: pTYB1 / Production host: Escherichia coli (E. coli) / Strain (production host): ER2566 / References: UniProt: D6E269
#2: RNA chain
RNA (45-MER)


Mass: 14556.560 Da / Num. of mol.: 8 / Source method: obtained synthetically / Source: (synth.) synthetic construct (others)
#3: Chemical...
ChemComp-CA / CALCIUM ION


Mass: 40.078 Da / Num. of mol.: 48 / Source method: obtained synthetically / Formula: Ca
#4: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 1322 / 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

Crystal
IDDensity Matthews3/Da)Density % sol (%)
13.0659.76
2
Crystal growTemperature: 293 K / Method: vapor diffusion / pH: 4.8
Details: CaCl2, 2-Propanol, Sodium Acetate, pH 4.8, vapor diffusion, temperature 293K

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Data collection

Diffraction
IDMean temperature (K)Crystal-ID
11001
21002
Diffraction source
SourceSiteBeamlineIDWavelength (Å)
SYNCHROTRONDiamond I0210.9795
SYNCHROTRONDiamond I0220.9795
Detector
TypeIDDetectorDate
DECTRIS PILATUS 6M1PIXELMay 8, 2013
DECTRIS PILATUS 6M2PIXELMay 8, 2013
Radiation
IDMonochromatorProtocolMonochromatic (M) / Laue (L)Scattering typeWavelength-ID
1Si 111 CHANNELSINGLE WAVELENGTHMx-ray1
2Si 111 CHANNELSINGLE WAVELENGTHMx-ray1
Radiation wavelengthWavelength: 0.9795 Å / Relative weight: 1
ReflectionRedundancy: 13.5 % / Number: 869571 / Rmerge(I) obs: 0.164 / D res high: 3 Å / D res low: 93.27 Å / Num. obs: 64337 / % possible obs: 100 / Rejects: 580
Diffraction reflection shell
Highest resolution (Å)Lowest resolution (Å)IDRmerge(I) obsRedundancy
33.0710.61212.6
13.7593.2710.05413.1
ReflectionResolution: 2.2→63.12 Å / Num. obs: 157960 / % possible obs: 97.9 % / Redundancy: 2.3 % / Rmerge(I) obs: 0.048 / Net I/σ(I): 11.6
Reflection shell
Resolution (Å)Redundancy (%)Rmerge(I) obsMean I/σ(I) obsDiffraction-ID% possible all
2.2-2.242.10.3382.41,297
12.05-63.042.60.02237.41,296.9

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Phasing

PhasingMethod: SAD

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Processing

Software
NameVersionClassificationNB
Aimless0.1.29data scaling
SOLVEphasing
REFMAC5.7.0029refinement
PDB_EXTRACT3.15data extraction
GDAdata collection
MOSFLMdata reduction
SCALAdata scaling
PHENIXphasing
RefinementMethod to determine structure: SAD / Resolution: 2.2→63.12 Å / Cor.coef. Fo:Fc: 0.952 / Cor.coef. Fo:Fc free: 0.938 / SU B: 5.894 / SU ML: 0.145 / SU R Cruickshank DPI: 0.2338 / Cross valid method: THROUGHOUT / ESU R: 0.234 / ESU R Free: 0.19 / Stereochemistry target values: MAXIMUM LIKELIHOOD / Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS
RfactorNum. reflection% reflectionSelection details
Rfree0.23386 7919 5 %RANDOM
Rwork0.19944 ---
obs0.2012 150001 97.85 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.2 Å / Solvent model: MASK
Displacement parametersBiso mean: 40.97 Å2
Baniso -1Baniso -2Baniso -3
1--2.65 Å20 Å20.57 Å2
2---0.23 Å20 Å2
3---2.84 Å2
Refinement stepCycle: LAST / Resolution: 2.2→63.12 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms10363 7696 48 1322 19429
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0130.01619265
X-RAY DIFFRACTIONr_bond_other_d0.0110.0213810
X-RAY DIFFRACTIONr_angle_refined_deg1.721.65327710
X-RAY DIFFRACTIONr_angle_other_deg1.854332321
X-RAY DIFFRACTIONr_dihedral_angle_1_deg6.10851248
X-RAY DIFFRACTIONr_dihedral_angle_2_deg34.26523.395542
X-RAY DIFFRACTIONr_dihedral_angle_3_deg17.686152040
X-RAY DIFFRACTIONr_dihedral_angle_4_deg22.6871587
X-RAY DIFFRACTIONr_chiral_restr0.1010.22903
X-RAY DIFFRACTIONr_gen_planes_refined0.0140.0216037
X-RAY DIFFRACTIONr_gen_planes_other0.0110.024378
X-RAY DIFFRACTIONr_nbd_refined
X-RAY DIFFRACTIONr_nbd_other
X-RAY DIFFRACTIONr_nbtor_refined
X-RAY DIFFRACTIONr_nbtor_other
X-RAY DIFFRACTIONr_xyhbond_nbd_refined
X-RAY DIFFRACTIONr_xyhbond_nbd_other
X-RAY DIFFRACTIONr_metal_ion_refined
X-RAY DIFFRACTIONr_metal_ion_other
X-RAY DIFFRACTIONr_symmetry_vdw_refined
X-RAY DIFFRACTIONr_symmetry_vdw_other
X-RAY DIFFRACTIONr_symmetry_hbond_refined
X-RAY DIFFRACTIONr_symmetry_hbond_other
X-RAY DIFFRACTIONr_symmetry_metal_ion_refined
X-RAY DIFFRACTIONr_symmetry_metal_ion_other
X-RAY DIFFRACTIONr_mcbond_it
X-RAY DIFFRACTIONr_mcbond_other
X-RAY DIFFRACTIONr_mcangle_it
X-RAY DIFFRACTIONr_mcangle_other
X-RAY DIFFRACTIONr_scbond_it
X-RAY DIFFRACTIONr_scbond_other
X-RAY DIFFRACTIONr_scangle_it
X-RAY DIFFRACTIONr_scangle_other
X-RAY DIFFRACTIONr_long_range_B_refined
X-RAY DIFFRACTIONr_long_range_B_other
X-RAY DIFFRACTIONr_rigid_bond_restr
X-RAY DIFFRACTIONr_sphericity_free
X-RAY DIFFRACTIONr_sphericity_bonded
Refine LS restraints NCS

Refine-ID: X-RAY DIFFRACTION / Type: interatomic distance / Weight position: 0.05

Ens-IDDom-IDAuth asym-IDNumberRms dev position (Å)
11A99190.09
12C99190.09
21A99240.09
22E99240.09
31A98800.09
32G98800.09
41A97810.08
42I97810.08
51A99020.1
52K99020.1
61A98860.08
62M98860.08
71A98340.1
72O98340.1
81B38720.06
82D38720.06
91B38790.06
92F38790.06
101B38780.05
102H38780.05
111B37860.09
112J37860.09
121B38660.05
122L38660.05
131B38370.08
132N38370.08
141B37500.12
142P37500.12
151C99050.09
152E99050.09
161C97520.1
162G97520.1
171C98120.08
172I98120.08
181C98500.1
182K98500.1
191C98710.09
192M98710.09
201C98560.09
202O98560.09
211D38720.04
212F38720.04
221D38770.03
222H38770.03
231D38010.07
232J38010.07
241D38550.04
242L38550.04
251D38180.09
252N38180.09
261D37460.11
262P37460.11
271E97190.1
272G97190.1
281E97290.09
282I97290.09
291E98110.1
292K98110.1
301E98390.09
302M98390.09
311E98210.1
312O98210.1
321F38780.03
322H38780.03
331F37720.08
332J37720.08
341F38760.03
342L38760.03
351F38200.09
352N38200.09
361F37540.11
362P37540.11
371G96780.09
372I96780.09
381G98450.1
382K98450.1
391G98270.09
392M98270.09
401G97430.1
402O97430.1
411H37740.08
412J37740.08
421H38620.03
422L38620.03
431H38320.08
432N38320.08
441H37650.11
442P37650.11
451I96940.1
452K96940.1
461I98350.08
462M98350.08
471I96780.09
472O96780.09
481J37690.08
482L37690.08
491J37660.1
492N37660.1
501J36900.13
502P36900.13
511K97790.1
512M97790.1
521K97300.11
522O97300.11
531L38090.09
532N38090.09
541L37410.11
542P37410.11
551M98130.09
552O98130.09
561N37340.13
562P37340.13
LS refinement shellResolution: 2.2→2.257 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.346 556 -
Rwork0.305 11102 -
obs--97.19 %

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