Extended Data Fig. 5: Active site titration of various BRAF variants to estimate active enzyme concentration using Morrison kinetics.
From: Negative regulation of RAF kinase activity by ATP is overcome by 14-3-3-induced dimerization
(Left) Active site titration of GDC-0879 against indicated enzymes. Activities for each enzyme were normalized to a no inhibitor control and fitted to Morrison Ki. For BRAFKD, low inhibitor concentration points below the observed paradoxical activation maximum activity were removed to allow for fitting of Morrison Ki. Each point represents three time points assayed in duplicate (n=6), normalized to reaction time. Error bars represent SD of six replicates. (Right) Zoomed version of active site titration of BRAFKD including all inhibitor points, showing paradoxical activation.
