8kal

Crystal structure of SpyCas9 in complex with sgRNA and 17nt target DNA

Method: X-RAY DIFFRACTION Dmax: 199.1 Å Quality: GOOD

1. Protein Identity and Related Structures Protein Identity & Related Structures

CRISPR-associated endonuclease Cas9/Csn1

Streptococcus pyogenes serotype M1

UniProt Q99ZW2

State in the Current Structure

Assembly Oligomeric State Construct Mutations and Modifications Ligands, Ions and Associated Components Method and Experimental Conditions Structure Quality
1 Other combination Monomer Protein × 1 DNA 2 RNA 1 PDB declaration: tetrameric(4) Consistent with all polymer counts Chain B; UniProt 1–1368 Not recorded RNA (98-MER) × 1 DNA (25-MER) × 1 ;DNA (5'-D(*TP*TP*TP*AP*GP*GP*TP*AP*TP*TP*G)-3') ; × 1 SO4 SULFATE ION × 1 X-RAY DIFFRACTION X-ray crystallization conditions:VAPOR DIFFUSION, HANGING DROP;289 K;0.2 M Ammonium sulfate, 0.1 M MES pH 6.0, 12.625% PEG8000 Resolution 3.16 Å R-free 0.288
2 Other combination Monomer Protein × 1 DNA 2 RNA 1 PDB declaration: tetrameric(4) Consistent with all polymer counts Chain G; UniProt 1–1368 Not recorded RNA (98-MER) × 1 DNA (25-MER) × 1 ;DNA (5'-D(*TP*TP*TP*AP*GP*GP*TP*AP*TP*TP*G)-3') ; × 1 SO4 SULFATE ION × 1 X-RAY DIFFRACTION X-ray crystallization conditions:VAPOR DIFFUSION, HANGING DROP;289 K;0.2 M Ammonium sulfate, 0.1 M MES pH 6.0, 12.625% PEG8000 Resolution 3.16 Å R-free 0.288

Other States of the Same Protein in the Database

Each row is a biological assembly of the same UniProt protein in another PDB entry. The “Difference from current entry” column identifies evidence-level differences; no tag means the currently parsed fields agree.

134 other PDB entries and 145 assemblies. Open the comparison page and filter oligomeric states

View Construct and Data Evidence
UniProt name CAS9_STRP1
Isoform
PDB entities 2
Chains and sequence ranges Author chain B; PDBConstruct 1–1368; UniProt 1–1368 Author chain G; PDBConstruct 1–1368; UniProt 1–1368

The page prioritizes protein identity, the current assembly, associated components, oligomeric state and cross-PDB links. Chain mapping and sequence ranges are retained as data evidence. Internal IDs, import timestamps and assembly operation expressions are maintenance fields and are not shown here.

SAXS scattering curve SAXS Profile

SAXS profile for 8kal

P(r) Distance Distribution P(r) Distribution

P(r) distribution for 8kal
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2. Structure Basics 2. Structure Basics

Entry ID entry_id8kal
Deposition date deposition_date2023-08-03
Structure title titleCrystal structure of SpyCas9 in complex with sgRNA and 17nt target DNA
Keywords keywordsNuclease, RNA BINDING PROTEIN/RNA/DNA, RNA BINDING PROTEIN-RNA-DNA complex; RNA BINDING PROTEIN/RNA/DNA
Experimental Method methodX-RAY DIFFRACTION

3. SAXS Parameters (CRYSOL theoretical calculation) 3. SAXS Parameters (CRYSOL)

Radius of gyration Rg (Guinier) rg_guinier62.28
Radius of gyration Rg (electron density) rg_electron62.16
Forward intensity I(0) i02795700000.00
Molecular weight molecular_weight390060.0 kDa
Excluded volume excluded_volume465850 ų
Envelope volume envelope_volume748140 ų
Hydration-shell volume shell_volume104330 ų
Envelope diameter envelope_diameter230.1
Shell Rg shell_rg59.75
Envelope Rg envelope_rg60.27
Shape Rg shape_rg62.13
Total Rg total_rg62.19
Total atoms total_atoms27151
Residues n_residues2913
Spherical-harmonic order n_harmonics20
q range q_range— – 0.5000 −1
Data points n_points101
Shell type shell_typedirectional
Solvent electron density solvent_density0.3340 e/ų
Shell contrast contrast_shell0.0300 e/ų
CRYSOL version crysol_version4.1.3

4. P(r) Distance Distribution (GNOM inversion) 4. P(r) Analysis (GNOM)

Maximum dimension Dmax dmax199.1
Rg (real space) rg_real62.79
Rg uncertainty (real space) rg_real_error1.48
I(0) (real space) i0_real2.7940e+09
I(0) uncertainty (real space) i0_real_error5.3840e+07
Rg (reciprocal space) rg_reciprocal61.71
I(0) (reciprocal space) i0_reciprocal2790000000.0000
Solution quality estimate total_estimate0.8125
Solution quality rating solution_quality GOOD a GOOD solution
P(r) peaks n_peaks2
Primary peak position r_peak_primary55.8
Skewness Skewness skewness0.494
Kurtosis Kurtosis kurtosis-0.466
Angular range angular_range— – 0.1250 −1
Current regularization parameter α current_alpha0.0013
Highest regularization parameter α highest_alpha261000000.0000
Real-space data points n_real_points26
GNOM version gnom_version4.1.3
Quality Criteria quality_criteria AN1: 0.000; Oscil: 0.828; Stabil: 0.999; Sysdev: 1.000; Positv: 1.000; Valcen: 0.983; Smooth: 0.092

5. Crystallography and Experiment 5. Crystallography & Experiment

6. Entities and Polymers Entities & Polymers (6)

8. Citations (1)

9. Files and Curves (10)