Restriction enzyme EcoRV catalytic rate (Kcat)

Value 0.9 min^-1
Organism Unspecified
Reference Halford SE, Goodall AJ. Modes of DNA cleavage by the EcoRV restriction endonuclease. Biochemistry. 1988 Mar 827(5):1771-7. p. 1775 right column 2nd paragraphPubMed ID2835097
Method To test the above hypothesis, that the mode of DNA cleavage by the EcoRV restriction enzyme is determined primarily by its fractional saturation with Mg2+, researchers used single turnover and transient kinetic methods to analyze DNA cleavage by EcoRV at both pH 7.5 and pH 6.0. Evaluation of k1 and k2 from single turnovers of EcoRV at pH 6.0 and 10 mM MgCl2 yielded average values of 1.7 min^-1 for k1 and 0.9 min^-1 for k2. For k1 and k2 see comments section
Comments DNA cleavage by the EcoRV restriction enzyme must involve the intermediates: (E1•E2)(S1•S2)?k1?(E1•E2)(P1•S2)?k2?(E1•E2)(P1•P2) where (E•E) is the protein dimer, (S•S) is the DNA duplex with an intact EcoRV recognition site, and (P•S) and (P•P) are DNA molecules cut at the EcoRV site in either one or both strands (eq. 1 p. 1775 left column).
Entered by Ben Marks
ID 101629