EcoRV
| Restriction endonuclease EcoRV | |||||||||
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EcoRV (pronounced "eco R five") is a type II restriction endonuclease isolated from certain strains of Escherichia coli. It has the alternative name Eco32I.
In
Structure
The structure of this enzyme, and several mutants, in complex with the DNA sequence which it cuts has been solved by X-ray crystallography.
The core of the enzyme consists of a five-stranded mixed β-sheet flanked by α-helices. The core is conserved in all other type II restriction endonucleases. It also has an N-terminal dimerization subdomain formed by a short α-helix, a two-stranded antiparallel -sheet, and a long α-helix. This subdomain is found only in EcoRV and PvuII.[2]
Mode of action
Like
Binding of the enzyme induces a conformational change in the DNA, bending it by about 50°. DNA bending results in the unstacking of the bases, widening of the minor groove, and compression of the major groove. This brings the phosphodiester linkage to be broken closer to the active site of the enzyme, where it can be cleaved. Cleavage occurs within the recognition sequence, and does not require ATP hydrolysis.[2]
EcoRV is the only type II restriction endonuclease known to cause a major protein-induced conformational change in the DNA.[2]
Uses
EcoRV is often used to cut open a
See also
- EcoRI, another nuclease enzyme from E. coli.
- EcoRII, another nuclease enzyme from E. coli.
- FokI, a nuclease enzyme from Flavobacterium okeanokoites[4]
