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. 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 7.

Steady-state kinetic parameters for hpol κ19-526 mis-insertion on G4 DNA and non-G4 DNA substrates prepared in LiCl

kcat (min−1) KM,dNTP (μM) kcat/KM,dNTP (min−1 μM−1) f ins a
18/42-mer: dCMP insertion opposite template dT (−5 nt from G22)
non-G4 DNA 0.13 ± 0.01 590 ± 100 0.00022 0.00016
G4 DNA 0.14 ± 0.01 580 ± 200 0.00024 0.00029
21/42-mer: dGMP insertion opposite template dG (−2 nt from G22)
non-G4 DNA 0.082 ± 0.005 190 ± 50 0.00043 0.0036
G4 DNA 0.10 ± 0.007 190 ± 50 0.00053 0.0048
23/42-mer: dGMP insertion opposite template dG (0 nt from G22)
non-G4 DNA 0.34 ± 0.02 570 ± 100 0.00060 0.0095
G4 DNA 0.31 ± 0.03 1300 ± 300 0.00024 0.0067
a

The mis-insertion frequency (fins) was calculated as (kcat/KM,incorrect dNTP)/(kcat/KM,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.