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PDBsum entry 5fq5

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protein dna_rna metals links
Hydrolase/DNA PDB id
5fq5
Jmol PyMol
Contents
Protein chain
1318 a.a.
DNA/RNA
Metals
__K ×13
_MG ×2
Waters ×1108
PDB id:
5fq5
Name: Hydrolase/DNA
Title: Crystal structure of cas9-sgrna-DNA complex solved by native sad phasing
Structure: Sgrna. Chain: a. Crispr-associated endonuclease cas9/csn1. Chain: b. Synonym: spycas9. Engineered: yes. Mutation: yes. Target DNA strand proximal fragment. Chain: c.
Source: Synthetic: yes. Synthetic construct. Organism_taxid: 32630. Streptococcus pyogenes. Organism_taxid: 1314. Gene: cas9, csn1, spy_1046. Expressed in: escherichia coli. Expression_system_taxid: 469008. Expression_system_variant: rosetta2.
Resolution:
2.14Å     R-factor:   0.191     R-free:   0.224
Authors: V.Olieric,T.Weinert,A.Finke,C.Anders,M.Jinek,M.Wang
Key ref: V.Olieric et al. (2016). Data-collection strategy for challenging native SAD phasing. Acta Crystallogr D Struct Biol, 72, 421-429. PubMed id: 26960129 DOI: 10.1107/S2059798315024110
Date:
07-Dec-15     Release date:   23-Mar-16    
PROCHECK
Go to PROCHECK summary
 Headers
 References

Protein chain
Pfam   ArchSchema ?
Q99ZW2  (Q99ZW2_STRP1) -  CRISPR-associated endonuclease Cas9/Csn1
Seq:
Struc:
 
Seq:
Struc:
 
Seq:
Struc:
1368 a.a.
1318 a.a.*
Key:    PfamA domain  Secondary structure
* PDB and UniProt seqs differ at 2 residue positions (black crosses)

 Enzyme reactions 
   Enzyme class: E.C.3.1.-.-
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]
 Gene Ontology (GO) functional annotation 
  GO annot!
  Biological process     nucleic acid phosphodiester bond hydrolysis   3 terms 
  Biochemical function     hydrolase activity     9 terms  

 

 
DOI no: 10.1107/S2059798315024110 Acta Crystallogr D Struct Biol 72:421-429 (2016)
PubMed id: 26960129  
 
 
Data-collection strategy for challenging native SAD phasing.
V.Olieric, T.Weinert, A.D.Finke, C.Anders, D.Li, N.Olieric, C.N.Borca, M.O.Steinmetz, M.Caffrey, M.Jinek, M.Wang.
 
  ABSTRACT  
 
Recent improvements in data-collection strategies have pushed the limits of native SAD (single-wavelength anomalous diffraction) phasing, a method that uses the weak anomalous signal of light elements naturally present in macromolecules. These involve the merging of multiple data sets from either multiple crystals or from a single crystal collected in multiple orientations at a low X-ray dose. Both approaches yield data of high multiplicity while minimizing radiation damage and systematic error, thus ensuring accurate measurements of the anomalous differences. Here, the combined use of these two strategies is described to solve cases of native SAD phasing that were particular challenges: the integral membrane diacylglycerol kinase (DgkA) with a low Bijvoet ratio of 1% and the large 200 kDa complex of the CRISPR-associated endonuclease (Cas9) bound to guide RNA and target DNA crystallized in the low-symmetry space group C2. The optimal native SAD data-collection strategy based on systematic measurements performed on the 266 kDa multiprotein/multiligand tubulin complex is discussed.
 

 

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