› 6k4q

X-ray diffraction
2.7Å resolution

Crystal structure of xCas9 in complex with sgRNA and DNA (CGG PAM)

Released:
DOI: 10.2210/pdb6k4q/pdb
PDB entry 6k4q coloured by chain, front view.
PDB entry 6k4q coloured by chain, side view.
PDB entry 6k4q coloured by chain, top view.
Source organisms:
Primary publication:
Molecular basis for the PAM expansion and fidelity enhancement of an evolved Cas9 nuclease.
Chen W, Zhang H, Zhang Y, Wang Y, Gan J, Ji Q
PLoS Biol 17 e3000496 (2019)
PMID: 31603896

Function and Biology Details

Biochemical function:
Biological process:
Cellular component:
  • not assigned
Sequence domains:

Structure analysis Details

Assembly composition:
hetero tetramer (preferred)
Assembly name:
PDBe Complex ID:
PDB-CPX-120122 (preferred)
Entry contents:
1 distinct polypeptide molecule
2 distinct DNA molecules
1 distinct RNA molecule
Macromolecules (4 distinct):
CRISPR-associated endonuclease Cas9/Csn1 Chain: B
Molecule details ›
Chain: B
Length: 1368 amino acids
Theoretical weight: 158.57 KDa
Source organism: Streptococcus pyogenes serotype M1
Expression system: Escherichia coli BL21(DE3)
UniProt:
  • Canonical: Q99ZW2 (Residues: 1-1368; Coverage: 100%)
Gene names: SPy_1046, cas9, csn1
Sequence domains:
DNA (28-MER) Chain: C
Molecule details ›
Chain: C
Length: 28 nucleotides
Theoretical weight: 8.57 KDa
Source organism: Streptococcus pyogenes
Expression system: Not provided
DNA (5'-D(*AP*AP*AP*CP*GP*GP*TP*AP*TP*TP*G)-3') Chain: D
Molecule details ›
Chain: D
Length: 12 nucleotides
Theoretical weight: 3.71 KDa
Source organism: Streptococcus pyogenes
Expression system: Not provided
sgRNA Chain: A
Molecule details ›
Chain: A
Length: 83 nucleotides
Theoretical weight: 26.79 KDa
Source organism: Streptococcus pyogenes
Expression system: Not provided

Ligands and Environments

1 bound ligand:
Ligand PO4 9 x PO4
No modified residues

Experiments and Validation Details

Entry percentile scores
X-ray source: SSRF BEAMLINE BL19U1
Spacegroup: C2
Unit cell:
a: 177.84Å b: 69.669Å c: 189.513Å
α: 90° β: 109.35° γ: 90°
R-values:
R R work R free
0.207 0.203 0.275
Expression systems:
  • Escherichia coli BL21(DE3)
  • Not provided

Quick links

Citations

5 review citations

Structural biology of CRISPR-Cas immunity and genome editing enzymes.
Wang et al. (2022)
4 more