DNA-tension dependence of restriction enzyme cleavage reveals details of the reaction pathway

DNA-tension dependence of restriction enzyme activity

The data show that the observed EcoRV cleavage rate on linearized pCco5 plasmid (number of recognition sites n = 1) decreases rapidly above of ∼30 pN DNA tension (enzyme concentration 25 nM in terms of dimers) ( Figure 5a). This decrease indicates that the tension induces another process in the cleavage reaction to become rate-limiting.

DNA-tension dependence of restriction enzyme activity

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The reaction pathway for orthodox type IIP restriction endonucleases. The association rate k on is the only rate that depends on enzyme concentration.

DNA-tension dependence of restriction enzyme activity

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In order to elucidate the connection between the mechanics and the chemistry of DNA recognition and cleavage, we used a single-molecule approach to measure rate changes in the reaction pathway of EcoRV and BamHI as a function of DNA tension.

DNA-tension dependence of restriction enzyme activity

DNA-tension dependence of restriction enzyme activity reveals mechanochemical properties of the reaction pathway. The restriction enzyme buffer for both EcoRV and BamHI contains 100 mM NaCl, 1 mM 2-mercaptoethanol, 5 mM MgCl2 and 10 mM Tris–HCl (pH 8.0). The data show that the observed EcoRV The force dependence of the cleavage

DNA-tension dependence of restriction enzyme cleavage

Title: DNA-tension dependence of restriction enzyme cleavage reveals details of the reaction pathway: Published in: Unknown journal, p.184A-184A. Author

Measured cleavage rate versus DNA tension. (a) The effe

DNA-tension dependence of restriction enzyme activity reveals mechanochemical properties of the reaction pathway.

DNA-tension dependence of restriction enzyme activity

DNA-tension dependence of restriction enzyme activity reveals mechanochemical properties of the reaction pathway

Dynamics of Single DNA Looping and Cleavage by Sau3AI and

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Dec 01, 2006 · Dependence of cleavage on DNA tension To quantify the effect of tension on cleavage, we chose an incubation time of 30 s so that the fraction of molecules cleaved was <100%. As the tension was increased from 0.06 to ∼0.7 pN, the activity decreased exponentially and the magnitude of the decrease was ∼10-fold at 0.7 pN ( Fig. 3 ).

Mechanism of DNA recognition by the restriction enzyme

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DNA-tension dependence of restriction enzyme activity reveals mechanochemical properties of the reaction pathway. van den Broek B. et al.

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Nucleic Acids Research, 2005, Vol. 33, No. 8 doi:10.1093

DNA-tension dependence of restriction enzyme activity reveals mechanochemical properties of the reaction pathway Bram van den Broek, Maarten C. Noom and Gijs J. L. Wuite*

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