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. Author manuscript; available in PMC: 2011 Mar 16.

Published in final edited form as: Biochemistry. 2010 Mar 16;49(10):2097–2109. doi: 10.1021/bi901977k

NS3h translocation kinetics monitored by stopped-flow experiments with fluorescein labeled ssDNA. NS3h (100 nM) was pre-incubated with 200 nM ssDNA labeled at the 5’ end with fluorescein (5’-F-(dT)_L at 37 °C in assay buffer and translocation was initiated by the addition of ATP:Mg²⁺ and heparin to a final concentration of 5 mM, 10 mM, and 4 mg/ml, respectively. Fluorescence time courses at short (A) and long (B) time scales obtained with 5’-F-(dT)₄₀ (), 5’-F-(dT)₆₀ (), 5’-F-(dT)₇₂ (), 5’-F-(dT)₈₈ () and 5’-F-(dT)₁₀₀ (). The solid lines in the figure are simulations using equation 1 and the best fit parameters obtained from the global NLLS analysis of the time courses using equation 1.

Inline graphic — NS3h translocation kinetics monitored by stopped-flow experiments with fluorescein labeled ssDNA. NS3h (100 nM) was pre-incubated with 200 nM ssDNA labeled at the 5’ end with fluorescein (5’-F-(dT)_L at 37 °C in assay buffer and translocation was initiated by the addition of ATP:Mg²⁺ and heparin to a final concentration of 5 mM, 10 mM, and 4 mg/ml, respectively. Fluorescence time courses at short (A) and long (B) time scales obtained with 5’-F-(dT)₄₀ (), 5’-F-(dT)₆₀ (), 5’-F-(dT)₇₂ (), 5’-F-(dT)₈₈ () and 5’-F-(dT)₁₀₀ (). The solid lines in the figure are simulations using equation 1 and the best fit parameters obtained from the global NLLS analysis of the time courses using equation 1.