Figure 5.
Michaelis–Menten saturation curve measured with the ADP‐Glo assay. MLC2v dose‐dependence of the nFlag‐tagged wild‐type or p.Pro639Valfs*15 cMLCK activities measured by ADP‐Glo assay. The reaction that contained no MLC2v was set as a background. The luminescence readings of the reactions were corrected with background subtraction and finally fit with the Michaelis–Menten equation using Igor Pro software to obtain Km (MLC2v). Wild‐type cMLCK had a Km value of 5.93 ± 1.47 μM and a maximum luminescence of (3.67 ± 0.37) × 104 relative light units that corresponded to a V max of 1.28 ± 0.03 mol/min/mol kinase. Mutant cMLCK had no kinase activity. n = 2 for each point. RLU, relative light unit.