Skip to main content
. 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 2.

Steady-state kinetic parameters for hpol κ19-526 catalysis on G4 DNA and non-G4 DNA substrates prepared in KCl

kcat (min−1) KM,dNTP (μM) kcat/KM,dNTP (min−1 μM−1) Relative activitya
13/42-mer: dCMP insertion opposite template dG −10 nt from G22
non-G4 DNA 7.5 ± 0.9 13 ± 3 0.58 -
G4 DNA 1.7 ± 0.2 30 ± 9 0.06 10.4 ± 1.4%
14/42-mer: dGMP insertion opposite template dC −9 nt from G22
non-G4 DNA 3.7 ± 1.0 26 ± 3 0.15 -
G4 DNA 1.2 ± 0.8 169 ± 37 0.007 5.0 ± 1.0%
18/42-mer: dAMP insertion opposite template dT −5 nt from G22
non-G4 DNA 8.9 ± 4.1 13 ± 2 0.65 -
G4 DNA 3.8 ± 0.8 85 ± 29 0.05 7.5 ± 2.1%
19/42-mer: dCMP insertion opposite template dG −4 nt from G22
non-G4 DNA 9.0 ± 3.5 20 ± 1 0.45 -
G4 DNA 2.3 ± 0.3 25 ± 8 0.10 22.8 ± 2.4%
20/42-mer: dGMP insertion opposite template dC −3 nt from G22
non-G4 DNA 5.1 ± 0.3 4.9 ± 1.5 1.10 -
G4 DNA 4.4 ± 1.0 4.1 ± 0.7 1.13 103 ± 18%
21/42-mer: dCMP insertion opposite template dG −2 nt from G22
non-G4 DNA 8.6 ± 1.8 28 ± 1 0.31 -
G4 DNA 7.9 ± 1.9 14 ± 2 0.57 188 ± 25%
22/42-mer: dAMP insertion opposite template dT −1 nt from G22
non-G4 DNA 7.3 ± 1.5 4.4 ± 0.9 1.67 -
G4 DNA 2.6 ± 0.4 11 ± 1 0.24 14.6 ± 3.1%
23/42-mer: dCMP insertion opposite G22
non-G4 DNA 6.4 ± 0.3 6.1 ± 0.4 1.06 -
G4 DNA 0.21 ± 0.03 26 ± 3 0.0081 0.76 ± 0.11%
a

Relative activity was calculated as [(kcat/Km,dNTP)G4 DNA/(kcat/Km,dNTP)non-G4 DNA] × 100 for three to four individual sets of experiments. For determination of kcat and Km,dNTP, 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 mean and standard deviation of three to four individual sets of experiments is reported for each kcat and Km,dNTP value.