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. 2014 Sep 27;42(19):12102–12111. doi: 10.1093/nar/gku879

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© The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 6. — Kinetic analysis of human PrimPol and PrimPol^Y89D. Single turnover kinetics of wild-type and PrimPol^Y89D to determine catalytic efficiency (k_pol) and dNTP binding efficiency (K_D(dNTP)). (A) The concentration of extended DNA product, as determined by electrophoresis, was plotted against time and fit to an exponential curve for a number of dATP concentrations for wild-type and PrimPol^Y89D. These data were fit to exponential curves as described in Equation (1) and k_obs was determined. (B) k_obs was subsequently plot against dATP concentration for wild-type PrimPol and PrimPol^Y89D and these data were fit to hyperbolic curves as described in Equation (2) to determine k_pol and K_D(dNTP). k_pol for wild-type PrimPol was found to be 6.98 min⁻¹ ± 0.40 and K_D(dNTP) was 15.51 μM ± 2.71. k_pol for PrimPol^Y89D was 4.07 min⁻¹ ± 0.19 and K_D(dNTP) was 170.21 μM ± 18.80.