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. 2015 Dec 10;10(12):e0144638. doi: 10.1371/journal.pone.0144638

Fig 2. Effect of the W501R substitution on ability of HCV helicase to unwind RNA.

Fig 2

(A) Representative fluorescence traces observed in split beacon helicase assay [36] withindicated amounts of each protein. The increase in rates of fluorescence intensity (RFU—Relative Fluorescence Unit) change indicates higher activity of NS3h_3a (wt) and NS3h_1b (con1) enzymes in RNA unwinding compared to the mutant protein. (B) Initial rates of unwinding after ATP addition in each reaction fit to the Michaelis-Menten equation. The reactions were prepared containing 25 mM MOPS, 1.25 mM MgCl2, 5 nM Cy5, 0.05 mM DTT, 0.005 mg/mL BSA, 0.001% Tween20, substrate, 1mM ATP and enzymes NS3h_1b (con1), NS3h_3a (wt) and NS3h_3a (W501R) at indicated concentrations.