Fig. 4
From: A continuous kinetic assay for protein and DNA methyltransferase enzymatic activities
Steady-state kinetic analysis of SETD6 activity with RelA protein (residues 1–431). a Kinetic traces of SETD6 at increased RelA concentrations of 0–2.5 μM. Initial rate at each RelA concentration was calculated by fitting the raw data to a linear equation. Absorbance values at 340 nm in all reactions are normalized to 1 and the change in absorbance over time is presented as 1—the absorbance value obtains for each measurement. b Fitting the initial rate data to linear equation allows the determination of k cat/K M to be 3.2·105 s−1 M−1. Background reaction was subtracted from initial rates. c Radioactive gel analysis of SETD6 activity with SAHN, ADE and RelA protein (1–431) showing that methylation does not take place on SAHN and ADE and only RelA is recognized as a substrate for SETD6