[English] 日本語
Yorodumi
- PDB-5u31: Crystal structure of AacC2c1-sgRNA-8mer substrate DNA ternary complex -

+
Open data


ID or keywords:

Loading...

-
Basic information

Entry
Database: PDB / ID: 5u31
TitleCrystal structure of AacC2c1-sgRNA-8mer substrate DNA ternary complex
Components
  • CRISPR-associated endonuclease C2c1
  • Non-target DNA strand
  • Target DNA strand
  • sgRNASubgenomic mRNA
KeywordsHYDROLASE/DNA / Type V CRISPR-Cas endonculease: C2c1: Structure: Binary complex with sgRNA: Ternary complex with added DNA: RuvC catalytic pocket: Sequence-specific PAM recognition: Genome editing tool / HYDROLASE-DNA complex
Function / homology
Function and homology information


defense response to virus / endonuclease activity / Hydrolases; Acting on ester bonds / DNA binding / RNA binding
Similarity search - Function
DNA / DNA (> 10) / RNA / RNA (> 10) / RNA (> 100) / CRISPR-associated endonuclease Cas12b
Similarity search - Component
Biological speciesAlicyclobacillus acidoterrestris (bacteria)
Homo sapiens (human)
MethodX-RAY DIFFRACTION / SYNCHROTRON / SAD / Resolution: 2.89 Å
AuthorsYang, H. / Gao, P. / Rajashankar, K.R. / Patel, D.J.
Funding support United States, 2items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)GM104962 United States
Memorial Sloan-Kettering Cancer Center Core GrantP30 CA008748 United States
CitationJournal: Cell / Year: 2016
Title: PAM-Dependent Target DNA Recognition and Cleavage by C2c1 CRISPR-Cas Endonuclease.
Authors: Yang, H. / Gao, P. / Rajashankar, K.R. / Patel, D.J.
History
DepositionDec 1, 2016Deposition site: RCSB / Processing site: RCSB
Revision 1.0Jan 25, 2017Provider: repository / Type: Initial release
Revision 1.1Feb 8, 2017Group: Structure summary
Revision 1.2Sep 20, 2017Group: Author supporting evidence / Category: pdbx_audit_support / Item: _pdbx_audit_support.funding_organization
Revision 1.3Jan 1, 2020Group: Author supporting evidence / Database references / Category: pdbx_audit_support / struct_ref_seq
Item: _pdbx_audit_support.funding_organization / _struct_ref_seq.db_align_beg / _struct_ref_seq.db_align_end

-
Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

-
Assembly

Deposited unit
A: CRISPR-associated endonuclease C2c1
B: sgRNA
C: Target DNA strand
D: Non-target DNA strand
E: Non-target DNA strand
hetero molecules


Theoretical massNumber of molelcules
Total (without water)181,5527
Polymers181,3605
Non-polymers1922
Water0
1


  • Idetical with deposited unit
  • defined by author&software
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area24220 Å2
ΔGint-205 kcal/mol
Surface area65660 Å2
MethodPISA
Unit cell
Length a, b, c (Å)117.249, 182.152, 216.789
Angle α, β, γ (deg.)90.00, 90.00, 90.00
Int Tables number23
Space group name H-MI222

-
Components

#1: Protein CRISPR-associated endonuclease C2c1 / AacC2c1


Mass: 131747.031 Da / Num. of mol.: 1 / Fragment: CRISPR-associated endonuclease AacC2c1 / Mutation: D570A/E848A/D977A
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Alicyclobacillus acidoterrestris (bacteria)
Strain: ATCC 49025 / DSM 3922 / CIP 106132 / NCIMB 13137 / GD3B
Gene: c2c1, N007_06525 / Plasmid: pRSFDuet-SUMO / Production host: Escherichia coli (E. coli) / Strain (production host): BL21(DE3)
References: UniProt: T0D7A2, Hydrolases; Acting on ester bonds
#2: RNA chain sgRNA / Subgenomic mRNA


Mass: 36183.555 Da / Num. of mol.: 1 / Source method: obtained synthetically
Source: (synth.) Alicyclobacillus acidoterrestris (bacteria)
#3: DNA chain Target DNA strand


Mass: 8532.490 Da / Num. of mol.: 1 / Source method: obtained synthetically / Source: (synth.) Homo sapiens (human)
#4: DNA chain Non-target DNA strand


Mass: 2448.613 Da / Num. of mol.: 2 / Source method: obtained synthetically / Source: (synth.) Homo sapiens (human)
#5: Chemical ChemComp-SO4 / SULFATE ION / Sulfate


Mass: 96.063 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: SO4

-
Experimental details

-
Experiment

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

-
Sample preparation

CrystalDensity Matthews: 3.2 Å3/Da / Density % sol: 61.52 %
Crystal growTemperature: 293 K / Method: evaporation / pH: 7.2
Details: 0.2 M ammonium acetate, 0.1 M HEPES (pH 7.5), and 25% PEG3350 (v/v)

-
Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 24-ID-E / Wavelength: 0.9792 Å
DetectorType: ADSC QUANTUM 315 / Detector: CCD / Date: Aug 22, 2016 / Details: LR-Design detector positioner
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.9792 Å / Relative weight: 1
ReflectionResolution: 2.89→50 Å / Num. obs: 52212 / % possible obs: 100 % / Redundancy: 14.3 % / Biso Wilson estimate: 59.26 Å2 / CC1/2: 0.996 / Net I/σ(I): 11.7
Reflection shellResolution: 2.89→2.98 Å / Redundancy: 11.8 % / Mean I/σ(I) obs: 1.2 / CC1/2: 0.48 / % possible all: 100

-
Processing

Software
NameVersionClassification
PHENIX(1.11.1_2575: ???)refinement
XDSdata scaling
PDB_EXTRACT3.22data extraction
XDSdata reduction
AutoSolphasing
SCALEPACKdata scaling
RefinementMethod to determine structure: SAD / Resolution: 2.89→49.295 Å / SU ML: 0.42 / Cross valid method: FREE R-VALUE / σ(F): 1.34 / Phase error: 25.74 / Stereochemistry target values: ML
RfactorNum. reflection% reflection
Rfree0.2393 2555 4.89 %
Rwork0.1976 --
obs0.1997 52212 99.96 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL
Refinement stepCycle: LAST / Resolution: 2.89→49.295 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms8864 2997 10 0 11871
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.00412415
X-RAY DIFFRACTIONf_angle_d0.6217413
X-RAY DIFFRACTIONf_dihedral_angle_d20.7597091
X-RAY DIFFRACTIONf_chiral_restr0.041925
X-RAY DIFFRACTIONf_plane_restr0.0041751
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
2.89-2.94560.37041360.33712715X-RAY DIFFRACTION100
2.9456-3.00570.31971480.30842748X-RAY DIFFRACTION100
3.0057-3.07110.40241160.29732703X-RAY DIFFRACTION100
3.0711-3.14250.32841500.29132751X-RAY DIFFRACTION100
3.1425-3.22110.33661500.28292686X-RAY DIFFRACTION100
3.2211-3.30810.34051340.24372729X-RAY DIFFRACTION100
3.3081-3.40550.27631480.22662751X-RAY DIFFRACTION100
3.4055-3.51540.25261340.21152733X-RAY DIFFRACTION100
3.5154-3.6410.25541490.20232748X-RAY DIFFRACTION100
3.641-3.78670.23481450.18712739X-RAY DIFFRACTION100
3.7867-3.9590.22581330.17682752X-RAY DIFFRACTION100
3.959-4.16760.20031470.1722765X-RAY DIFFRACTION100
4.1676-4.42850.19141340.1562750X-RAY DIFFRACTION100
4.4285-4.77020.19041370.15222773X-RAY DIFFRACTION100
4.7702-5.24980.21171380.16262792X-RAY DIFFRACTION100
5.2498-6.00830.21491300.17262795X-RAY DIFFRACTION100
6.0083-7.56540.2311620.19162802X-RAY DIFFRACTION100
7.5654-49.30220.20631640.18582925X-RAY DIFFRACTION100
Refinement TLS params.Method: refined / Origin x: 145.0204 Å / Origin y: 207.3176 Å / Origin z: 284.3392 Å
111213212223313233
T0.4264 Å2-0.0295 Å2-0.0082 Å2-0.3447 Å20.0315 Å2--0.3479 Å2
L1.0401 °20.1875 °2-0.3648 °2-0.5221 °2-0.2095 °2--0.5083 °2
S-0.0215 Å °0.0776 Å °-0.1136 Å °-0.0577 Å °-0.0431 Å °-0.1892 Å °-0.0112 Å °0.1114 Å °0.0589 Å °
Refinement TLS groupSelection details: all

+
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