. Author manuscript; available in PMC: 2017 Sep 20.

Published in final edited form as: Biochemistry. 2016 Sep 7;55(37):5218–5229. doi: 10.1021/acs.biochem.6b00374

Table 5.

Steady-state kinetic parameters for hpol κ^19-526 mis-insertion on G4 DNA and non-G4 DNA substrates prepared in KCl

	k_cat (min⁻¹)	K_M,dNTP (μM)	k_cat/K_M,dNTP (min⁻¹ μM⁻¹)	f _ins ^a
18/42-mer: dCMP insertion opposite template dT (−5 nt from G22)
non-G4 DNA	1.3 ± 0.3	1500 ± 570	0.00087	0.00087
G4 DNA	0.14 ± 0.05	1600 ± 860	0.000088	0.0018
21/42-mer: dGMP insertion opposite template dG (−2 nt from G22)
non-G4 DNA	0.21 ± 0.01	350 ± 60	0.00060	0.0018
G4 DNA	0.42 ± 0.02	460 ± 70	0.00091	0.0014
21/42-mer: dAMP insertion opposite template dG (−2 nt from G22)
non-G4 DNA	0.069 ± 0.01	1400 ± 500	0.000049	0.00014
G4 DNA	0.28 ± 0.01	350 ± 50	0.00080	0.0013
23/42-mer: dGMP insertion opposite template dG (0 nt from G22)
non-G4 DNA	2.2 ± 0.3	1100 ± 300	0.0020	0.0056
G4 DNA	0.0053 ± 0.0007	420 ± 200	0.000013	0.00062

The mis-insertion frequency (f_ins) was calculated as (k_cat/K_{M,incorrect dNTP})/(k_cat/K_{M,correct dNTP}). For determination of kinetic values, graphs of the initial rate of product formation versus dNTP concentration were plotted using non-linear regression analysis (one-site hyperbolic fit) in the GraphPad Prism program. The standard error of the fit is reported for each value.