[English] 日本語
Yorodumi
- PDB-5b2q: Crystal structure of Francisella novicida Cas9 RHA in complex wit... -

+
Open data


ID or keywords:

Loading...

-
Basic information

Entry
Database: PDB / ID: 5b2q
TitleCrystal structure of Francisella novicida Cas9 RHA in complex with sgRNA and target DNA (TGG PAM)
Components
  • CRISPR-associated endonuclease Cas9
  • DNA (5'-D(*TP*GP*GP*TP*AP*TP*CP*GP*G)-3')
  • Guide RNA
  • Target DNA
KeywordsHYDROLASE/RNA/DNA / CRISPR-Cas9 / genome engineering / HYDROLASE-RNA-DNA complex
Function / homology
Function and homology information


defense response to virus / endonuclease activity / Hydrolases; Acting on ester bonds / DNA binding / RNA binding / metal ion binding
Similarity search - Function
CRISPR system subtype II-B RNA-guided endonuclease Cas9/Csx12 / : / CRISPR-associated endonuclease Cas9 alpha-helical lobe / CRISPR-associated endonuclease Cas9, C-terminal domain / Cas9-type HNH domain / Cas9-type HNH domain profile.
Similarity search - Domain/homology
ACETATE ION / DNA / DNA (> 10) / RNA / RNA (> 10) / CRISPR-associated endonuclease Cas9
Similarity search - Component
Biological speciesFrancisella tularensis subsp. novicida U112 (bacteria)
synthetic construct (others)
Francisella tularensis subsp. novicida (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / SAD / Resolution: 1.7 Å
AuthorsHirano, H. / Nishimasu, H. / Nakane, T. / Ishitani, R. / Nureki, O.
CitationJournal: Cell / Year: 2016
Title: Structure and Engineering of Francisella novicida Cas9
Authors: Hirano, H. / Gootenberg, J.S. / Horii, T. / Abudayyeh, O.O. / Kimura, M. / Hsu, P.D. / Nakane, T. / Ishitani, R. / Hatada, I. / Zhang, F. / Nishimasu, H. / Nureki, O.
History
DepositionFeb 1, 2016Deposition site: PDBJ / Processing site: PDBJ
Revision 1.0Mar 2, 2016Provider: repository / Type: Initial release
Revision 1.1Mar 9, 2016Group: Database references
Revision 1.2Oct 4, 2017Group: Advisory / Data collection / Derived calculations
Category: diffrn_detector / diffrn_source ...diffrn_detector / diffrn_source / pdbx_struct_oper_list / pdbx_unobs_or_zero_occ_atoms
Item: _diffrn_detector.detector / _diffrn_source.pdbx_synchrotron_site / _pdbx_struct_oper_list.symmetry_operation
Revision 1.3Mar 20, 2024Group: Data collection / Database references / Derived calculations
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / pdbx_struct_conn_angle / struct_conn / struct_conn_type
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession ..._database_2.pdbx_DOI / _database_2.pdbx_database_accession / _pdbx_struct_conn_angle.ptnr1_auth_asym_id / _pdbx_struct_conn_angle.ptnr1_auth_comp_id / _pdbx_struct_conn_angle.ptnr1_auth_seq_id / _pdbx_struct_conn_angle.ptnr1_label_asym_id / _pdbx_struct_conn_angle.ptnr1_label_atom_id / _pdbx_struct_conn_angle.ptnr1_label_comp_id / _pdbx_struct_conn_angle.ptnr1_label_seq_id / _pdbx_struct_conn_angle.ptnr2_auth_asym_id / _pdbx_struct_conn_angle.ptnr2_auth_comp_id / _pdbx_struct_conn_angle.ptnr2_auth_seq_id / _pdbx_struct_conn_angle.ptnr2_label_asym_id / _pdbx_struct_conn_angle.ptnr2_label_atom_id / _pdbx_struct_conn_angle.ptnr2_label_comp_id / _pdbx_struct_conn_angle.ptnr3_auth_asym_id / _pdbx_struct_conn_angle.ptnr3_auth_comp_id / _pdbx_struct_conn_angle.ptnr3_auth_seq_id / _pdbx_struct_conn_angle.ptnr3_label_asym_id / _pdbx_struct_conn_angle.ptnr3_label_atom_id / _pdbx_struct_conn_angle.ptnr3_label_comp_id / _pdbx_struct_conn_angle.ptnr3_label_seq_id / _pdbx_struct_conn_angle.value / _struct_conn.conn_type_id / _struct_conn.id / _struct_conn.pdbx_dist_value / _struct_conn.pdbx_leaving_atom_flag / _struct_conn.pdbx_ptnr1_label_alt_id / _struct_conn.pdbx_value_order / _struct_conn.ptnr1_auth_asym_id / _struct_conn.ptnr1_auth_comp_id / _struct_conn.ptnr1_auth_seq_id / _struct_conn.ptnr1_label_asym_id / _struct_conn.ptnr1_label_atom_id / _struct_conn.ptnr1_label_comp_id / _struct_conn.ptnr1_label_seq_id / _struct_conn.ptnr2_auth_asym_id / _struct_conn.ptnr2_auth_comp_id / _struct_conn.ptnr2_auth_seq_id / _struct_conn.ptnr2_label_asym_id / _struct_conn.ptnr2_label_atom_id / _struct_conn.ptnr2_label_comp_id / _struct_conn.ptnr2_label_seq_id / _struct_conn_type.id

-
Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

-
Assembly

Deposited unit
A: CRISPR-associated endonuclease Cas9
B: Guide RNA
C: Target DNA
D: DNA (5'-D(*TP*GP*GP*TP*AP*TP*CP*GP*G)-3')
hetero molecules


Theoretical massNumber of molelcules
Total (without water)235,52354
Polymers233,0064
Non-polymers2,51750
Water17,889993
1


  • Idetical with deposited unit
  • defined by author&software
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area32640 Å2
ΔGint-333 kcal/mol
Surface area77190 Å2
MethodPISA
Unit cell
Length a, b, c (Å)81.562, 158.977, 96.738
Angle α, β, γ (deg.)90.000, 106.880, 90.000
Int Tables number4
Space group name H-MP1211

-
Components

-
DNA chain , 2 types, 2 molecules CD

#3: DNA chain Target DNA


Mass: 8993.800 Da / Num. of mol.: 1 / Source method: obtained synthetically / Details: chemical synthesis
Source: (synth.) Francisella tularensis subsp. novicida (bacteria)
#4: DNA chain DNA (5'-D(*TP*GP*GP*TP*AP*TP*CP*GP*G)-3')


Mass: 2786.833 Da / Num. of mol.: 1 / Source method: obtained synthetically
Source: (synth.) Francisella tularensis subsp. novicida (bacteria)

-
Protein / RNA chain , 2 types, 2 molecules AB

#1: Protein CRISPR-associated endonuclease Cas9


Mass: 190946.422 Da / Num. of mol.: 1 / Mutation: N995A
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Francisella tularensis subsp. novicida U112 (bacteria)
Strain: U112 / Gene: cas9, FTN_0757 / Plasmid: pE-SUMO / Production host: Escherichia coli (E. coli) / Strain (production host): BL21(DE3) Rosetta2
References: UniProt: A0Q5Y3, Hydrolases; Acting on ester bonds
#2: RNA chain Guide RNA


Mass: 30278.863 Da / Num. of mol.: 1 / Source method: obtained synthetically / Details: in vitro transcription / Source: (synth.) synthetic construct (others)

-
Non-polymers , 7 types, 1043 molecules

#5: Chemical ChemComp-ZN / ZINC ION


Mass: 65.409 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: Zn
#6: Chemical
ChemComp-NA / SODIUM ION


Mass: 22.990 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: Na
#7: Chemical ChemComp-CL / CHLORIDE ION


Mass: 35.453 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: Cl
#8: Chemical
ChemComp-CA / CALCIUM ION


Mass: 40.078 Da / Num. of mol.: 17 / Source method: obtained synthetically / Formula: Ca
#9: Chemical...
ChemComp-EDO / 1,2-ETHANEDIOL / ETHYLENE GLYCOL


Mass: 62.068 Da / Num. of mol.: 24 / Source method: obtained synthetically / Formula: C2H6O2
#10: Chemical ChemComp-ACT / ACETATE ION


Mass: 59.044 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: C2H3O2
#11: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 993 / Source method: isolated from a natural source / Formula: H2O

-
Experimental details

-
Experiment

ExperimentMethod: X-RAY DIFFRACTION / Number of used crystals: 1

-
Sample preparation

CrystalDensity Matthews: 2.58 Å3/Da / Density % sol: 52.25 %
Crystal growTemperature: 293 K / Method: vapor diffusion, hanging drop / pH: 5
Details: 9-11% PEG 3350, 0.2 M calcium acetate, 0.1 M sodium acetate

-
Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: SPring-8 / Beamline: BL41XU / Wavelength: 1 Å
DetectorType: DECTRIS PILATUS 6M / Detector: PIXEL / Date: Jun 29, 2015
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 1 Å / Relative weight: 1
ReflectionResolution: 1.7→46.29 Å / Num. obs: 254994 / % possible obs: 98.9 % / Redundancy: 7 % / Net I/σ(I): 20.3

-
Processing

Software
NameVersionClassification
Aimlessdata scaling
PHENIX1.10_2155: ???refinement
PDB_EXTRACT3.2data extraction
RefinementMethod to determine structure: SAD / Resolution: 1.7→46.286 Å / SU ML: 0.21 / Cross valid method: FREE R-VALUE / σ(F): 1.38 / Phase error: 21.55
RfactorNum. reflection% reflection
Rfree0.206 12762 5.01 %
Rwork0.1831 --
obs0.1842 254945 98.88 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å
Displacement parametersBiso max: 147.59 Å2 / Biso mean: 48.8681 Å2 / Biso min: 13.86 Å2
Refinement stepCycle: final / Resolution: 1.7→46.286 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms11701 2771 128 993 15593
Biso mean--49.29 40.82 -
Num. residues----1588
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.00815216
X-RAY DIFFRACTIONf_angle_d0.99621129
X-RAY DIFFRACTIONf_chiral_restr0.0562415
X-RAY DIFFRACTIONf_plane_restr0.0062232
X-RAY DIFFRACTIONf_dihedral_angle_d16.9438857
LS refinement shell

Refine-ID: X-RAY DIFFRACTION / Total num. of bins used: 30

Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkNum. reflection all% reflection obs (%)
1.7-1.71930.34044280.32947955838399
1.7193-1.73960.34894100.31598076848699
1.7396-1.76080.31774300.2998050848099
1.7608-1.78310.2874310.28618033846499
1.7831-1.80650.27954210.26818106852799
1.8065-1.83130.26414270.2647990841799
1.8313-1.85740.28074230.24728096851999
1.8574-1.88520.27824290.24627985841499
1.8852-1.91460.23114210.2247972839398
1.9146-1.9460.24484200.22027956837697
1.946-1.97960.22814250.20628045847099
1.9796-2.01560.23554210.20098101852299
2.0156-2.05430.24044240.20428096852099
2.0543-2.09630.2164300.19218102853299
2.0963-2.14180.22574220.19268131855399
2.1418-2.19170.20964270.17878046847399
2.1917-2.24650.20554230.1768074849799
2.2465-2.30720.19884220.17898043846598
2.3072-2.37510.20574190.1797997841698
2.3751-2.45180.21444280.1798111853999
2.4518-2.53940.21554270.174881328559100
2.5394-2.6410.20514300.17848101853199
2.641-2.76120.23484270.18788100852799
2.7612-2.90680.20344340.18128159859399
2.9068-3.08890.1924130.1757956836998
3.0889-3.32730.18714280.170881588586100
3.3273-3.6620.20614260.169581428568100
3.662-4.19160.1824340.1548140857499
4.1916-5.27980.16794280.15048098852698
5.2798-46.30310.18514340.18438232866699
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)
11.3623-0.2191.21830.5685-0.15244.1167-0.0516-0.23360.1810.13050.0636-0.0508-0.1347-0.0760.04810.1423-00.01930.1418-0.06460.19343.94412.440425.2416
21.65742.2008-0.88291.7856-1.0434-0.1896-0.13930.2893-1.1175-0.29740.0343-0.05340.4619-0.0740.0920.6853-0.08620.0150.3767-0.12421.1641-15.9766-33.79570.1748
31.5476-0.39821.33740.4061-0.16951.9260.04880.32860.0650.0222-0.0279-0.08760.15320.5552-0.02410.17810.01380.0460.306-0.03510.215327.0186-8.30722.6813
41.03170.4030.32513.99941.44460.8527-0.1037-0.0989-0.38290.61560.27620.22210.54890.1583-0.14740.54490.05690.03380.33290.00590.434422.8719-31.413528.0989
51.508-0.64410.03321.20420.41111.3222-0.0646-0.2598-0.21770.18170.14710.03930.36970.0342-0.07010.37890.0270.00010.3130.01880.230918.0581-19.952138.5635
61.9525-0.9211-0.10481.6797-1.28651.59710.5185-0.0593-0.4536-0.4267-0.28-0.05610.84160.7184-0.28841.16580.0406-0.0731.07020.29130.81027.8485-30.776565.1718
72.60640.0899-0.29113.1877-0.42122.6023-0.0484-0.9657-0.34340.4034-0.0241-0.00530.3276-0.25290.04540.3246-0.0056-0.02830.62090.01920.20859.5123-9.155656.0308
81.2541-0.38480.65270.6592-0.23061.0305-0.1251-0.1830.13550.02950.05550.0849-0.2174-0.20010.04640.2320.00750.03040.1661-0.03120.2644-22.077911.95511.6056
91.1935-0.01870.21052.7767-0.15042.1929-0.1359-0.53330.40671.14660.14060.2058-0.6996-0.2191-0.00580.7430.16760.0360.3859-0.16610.3398-15.93519.244535.9444
103.0051-0.36881.07291.43960.08781.81780.1686-0.6456-0.0960.15840.02790.15010.45340.1474-0.30440.7030.08470.04880.4820.04180.503316.6456-24.434637.1521
111.973-0.73921.59960.6092-0.42183.05890.0659-0.0817-0.1714-0.01520.03590.17590.2639-0.0133-0.1560.1954-0.00230.02330.1654-0.0090.22462.5605-9.901317.6563
122.143-0.59380.68051.3274-0.4372.79180.08590.2206-0.0806-0.0665-0.05010.1269-0.02040.04360.00050.2004-0.0131-0.00180.1701-0.01230.2287-16.45153.1365-2.5431
135.0471-1.13160.81344.1089-1.12623.640.06890.77730.1782-0.6722-0.07620.5025-0.31310.0475-0.02280.41310.0286-0.14030.4174-0.05730.3703-34.05938.2659-16.0582
141.6274-0.66790.60510.585-0.12861.0276-0.00380.12460.0341-0.0835-0.03350.0901-0.04320.01710.04230.205-0.0171-0.00090.12890.01460.2141-15.61896.288-0.0879
154.78210.0787-3.4110.03040.10553.39960.2705-0.05360.93760.2544-0.5725-0.1997-0.90660.17810.24920.3799-0.11680.00080.30120.02760.42877.822913.46868.723
161.8133-0.4646-0.22410.17140.02151.3505-0.1683-0.10130.179-0.06970.0311-0.0251-0.11730.12410.1420.2136-0.0082-0.00060.1892-0.0390.26152.41986.961322.3619
178.10225.0358-1.2676.8191-4.48015.9159-0.0445-0.48170.81530.7234-0.0577-0.2356-0.7749-0.49660.07230.2350.0778-0.07170.2835-0.07830.22929.52147.971330.294
183.9768-1.3830.62110.7583-0.06780.2843-0.00940.56440.5734-0.0152-0.1435-0.3061-0.03810.42720.13220.1696-0.03350.0180.39270.03210.283225.4620.416815.8895
193.20393.1994-3.39684.0176-3.27043.6436-0.12790.9664-0.6629-1.1009-0.2422-0.74171.21630.5390.34460.41260.13220.07880.6074-0.12010.341227.3654-14.48379.1201
203.81681.08660.10642.7463-2.16512.3916-0.08840.32450.62270.1017-0.1406-0.2228-0.10030.37010.19670.1259-0.0332-0.01010.35420.02350.318125.2048-0.333216.2347
215.667-2.5277-2.86245.61661.50652.4145-0.4282-0.02060.20540.6728-0.08011.2526-0.3005-1.33480.37850.40710.27280.0240.7108-0.05990.5938-27.522513.199328.0354
220.7748-0.25320.22890.82710.06481.1142-0.1154-0.424-0.48410.44340.19270.15880.21410.1252-0.15980.45490.09190.07150.34460.11440.325510.0414-20.530929.2929
234.4722-1.1304-0.93561.62350.68670.37-0.23240.10430.77530.30950.20271.27-0.7159-1.2440.13590.44950.32880.04670.86570.00160.8371-28.650416.552127.1303
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection detailsAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1chain 'A' and (resid 1 through 99 )A1 - 99
2X-RAY DIFFRACTION2chain 'A' and (resid 100 through 324 )A100 - 324
3X-RAY DIFFRACTION3chain 'A' and (resid 325 through 643 )A325 - 643
4X-RAY DIFFRACTION4chain 'A' and (resid 644 through 750 )A644 - 750
5X-RAY DIFFRACTION5chain 'A' and (resid 751 through 927 )A751 - 927
6X-RAY DIFFRACTION6chain 'A' and (resid 928 through 1065 )A928 - 1065
7X-RAY DIFFRACTION7chain 'A' and (resid 1066 through 1176 )A0
8X-RAY DIFFRACTION8chain 'A' and (resid 1177 through 1457 )A0
9X-RAY DIFFRACTION9chain 'A' and (resid 1458 through 1622 )A0
10X-RAY DIFFRACTION10chain 'B' and (resid 1 through 10 )B1 - 10
11X-RAY DIFFRACTION11chain 'B' and (resid 11 through 20 )B11 - 20
12X-RAY DIFFRACTION12chain 'B' and (resid 21 through 30 )B21 - 30
13X-RAY DIFFRACTION13chain 'B' and (resid 31 through 40 )B31 - 40
14X-RAY DIFFRACTION14chain 'B' and (resid 41 through 55 )B41 - 55
15X-RAY DIFFRACTION15chain 'B' and (resid 56 through 60 )B56 - 60
16X-RAY DIFFRACTION16chain 'B' and (resid 61 through 65 )B61 - 65
17X-RAY DIFFRACTION17chain 'B' and (resid 66 through 70 )B66 - 70
18X-RAY DIFFRACTION18chain 'B' and (resid 71 through 80 )B71 - 80
19X-RAY DIFFRACTION19chain 'B' and (resid 81 through 85 )B81 - 85
20X-RAY DIFFRACTION20chain 'B' and (resid 86 through 94 )B86 - 94
21X-RAY DIFFRACTION21chain 'C' and (resid 1 through 10 )C1 - 10
22X-RAY DIFFRACTION22chain 'C' and (resid 11 through 30 )C11 - 30
23X-RAY DIFFRACTION23chain 'D' and (resid 1 through 9 )D1 - 9

+
About Yorodumi

-
News

-
Feb 9, 2022. New format data for meta-information of EMDB entries

New format data for meta-information of EMDB entries

  • Version 3 of the EMDB header file is now the official format.
  • The previous official version 1.9 will be removed from the archive.

Related info.:EMDB header

External links:wwPDB to switch to version 3 of the EMDB data model

-
Aug 12, 2020. Covid-19 info

Covid-19 info

URL: https://pdbj.org/emnavi/covid19.php

New page: Covid-19 featured information page in EM Navigator.

Related info.:Covid-19 info / Mar 5, 2020. Novel coronavirus structure data

+
Mar 5, 2020. Novel coronavirus structure data

Novel coronavirus structure data

Related info.:Yorodumi Speices / Aug 12, 2020. Covid-19 info

External links:COVID-19 featured content - PDBj / Molecule of the Month (242):Coronavirus Proteases

+
Jan 31, 2019. EMDB accession codes are about to change! (news from PDBe EMDB page)

EMDB accession codes are about to change! (news from PDBe EMDB page)

  • The allocation of 4 digits for EMDB accession codes will soon come to an end. Whilst these codes will remain in use, new EMDB accession codes will include an additional digit and will expand incrementally as the available range of codes is exhausted. The current 4-digit format prefixed with “EMD-” (i.e. EMD-XXXX) will advance to a 5-digit format (i.e. EMD-XXXXX), and so on. It is currently estimated that the 4-digit codes will be depleted around Spring 2019, at which point the 5-digit format will come into force.
  • The EM Navigator/Yorodumi systems omit the EMD- prefix.

Related info.:Q: What is EMD? / ID/Accession-code notation in Yorodumi/EM Navigator

External links:EMDB Accession Codes are Changing Soon! / Contact to PDBj

+
Jul 12, 2017. Major update of PDB

Major update of PDB

  • wwPDB released updated PDB data conforming to the new PDBx/mmCIF dictionary.
  • This is a major update changing the version number from 4 to 5, and with Remediation, in which all the entries are updated.
  • In this update, many items about electron microscopy experimental information are reorganized (e.g. em_software).
  • Now, EM Navigator and Yorodumi are based on the updated data.

External links:wwPDB Remediation / Enriched Model Files Conforming to OneDep Data Standards Now Available in the PDB FTP Archive

-
Yorodumi

Thousand views of thousand structures

  • Yorodumi is a browser for structure data from EMDB, PDB, SASBDB, etc.
  • This page is also the successor to EM Navigator detail page, and also detail information page/front-end page for Omokage search.
  • The word "yorodu" (or yorozu) is an old Japanese word meaning "ten thousand". "mi" (miru) is to see.

Related info.:EMDB / PDB / SASBDB / Comparison of 3 databanks / Yorodumi Search / Aug 31, 2016. New EM Navigator & Yorodumi / Yorodumi Papers / Jmol/JSmol / Function and homology information / Changes in new EM Navigator and Yorodumi

Read more