RAS ubiquitylation modulates effector interactions

Figures & data

Table 1. Comparative binding affinity of wt KRAS and mUbRas with GAP and effector proteins.

	CRAF	RalGDS	PI3Kcγ	GAPcat	CRAF (GDP)
wt KRAS	61.0 ± 0.53 nM	1.01 ± 0.08 μM	1.86 ± 0.29 μM	1.9 ± 0.20 μM	55.2 ± 6.1 μM
mUbRAS	391 ± 21.20 nM	7.14 ± 0.51 μM	18.9 ± 2.02 μM	19.4 ± 1.4 μM	1.2 ± 0.2 μM

Figure 1. Panel A: Fluorescence binding curve of KRAS (mGppCp) with GAPcat (K_d = 1.9 ± 0.2 μM) compared with Panel B: Fluorescence binding assay of mUbRAS (mGppCp) with GAPcat (K_d = 19.4± 1.4 μM). Panel C: Relative binding affinity of KRAS (mGppCp, black bars) and mUbRAS (mGppCp, blue bars) with effectors CRAF RBD, RALGDS, and PI3Kcγ. Monoubiquitylated KRAS (mGppCp) shows a 7 to 9-decrease in affinity to the CRAF RBD, RalGDS RBD, and PI3Kcγ. Statistical error was determined from 3–5 independent experiments. Panel D: In contrast, mUbRAS (GDP shows 45-fold higher affinity relative to wt KRAS (GDP). Statistical error was determined from 3–5 independent experiments.