. 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 6.

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

	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	1.6 ± 0.4	1400 ± 500	0.0011	0.0014
G4 DNA	1.1 ± 0.3	2600 ± 800	0.00042	0.0012
21/42-mer: dGMP insertion opposite template dG (−2 nt from G22)
non-G4 DNA	0.12 ± 0.004	230 ± 30	0.00052	0.0025
G4 DNA	0.16 ± 0.007	290 ± 40	0.00056	0.0019
21/42-mer: dAMP insertion opposite template dG (−2 nt from G22)
non-G4 DNA	0.073 ± 0.007	1200 ± 300	0.000061	0.00029
G4 DNA	0.24 ± 0.02	430 ± 90	0.00056	0.0019
23/42-mer: dGMP insertion opposite template dG (0 nt from G22)
non-G4 DNA	2.0 ± 0.1	770 ± 100	0.0026	0.0020
G4 DNA	0.62 ± 0.09	2600 ± 600	0.00024	0.0017

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.