FokI
| Restriction endonuclease Fok1, C terminal | |||||||||
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The restriction endonuclease Fok1, naturally found in
Its molecular mass is 65.4 kDa, being composed of 587 amino acids.
DNA-binding domain
The recognition domain contains three subdomains (D1, D2 and D3) that are evolutionarily related to the DNA-binding domain of the catabolite gene activator protein which contains a helix-turn-helix.[3]
DNA-cleavage domain
DNA cleavage is mediated through the non-specific cleavage domain which also includes the dimerisation surface.[4] The dimer interface is formed by the parallel helices α4 and α5 and two loops P1 and P2 of the cleavage domain.[3]
Activity
When the nuclease is unbound to DNA, the endonuclease domain is sequestered by the DNA-binding domain and is released through a conformational change in the DNA-binding domain upon binding to its recognition site. Cleavage only occurs upon dimerization, when the recognition domain is bound to its cognate site and in the presence of magnesium ions.[4]
Exploitation
The endonuclease domain of Fok1 has been used in several studies, after combination with a variety of
One of several human
References
- S2CID 205027830.
- ^ PMID 16251401.
- ^ PMID 9724743.
- ^ PMID 9724744.
- doi:10.1038/nbt.2908
- doi:10.1038/nbt.2909
- S2CID 22436824.
See also
- Protein engineering
- Genetic engineering
- Zinc finger nuclease