Extended Data Fig. 7: SHOC2:PP1C:RAS interaction data.
From: Structural basis for SHOC2 modulation of RAS signalling
a, SEC trace of complex formation between SHOC2ΔN, PP1C and KRAS(GCP) (orange) (top) with associated SDS-PAGE analysis of fractions (bottom). b, Peptide dephosphorylation showing KDapp for SHOC2 and SHOC2ΔN. [PP1C] = 25 nM, [MRAS(GCP)] = 1 µM. Data points are from two time points (n = 2) (left). Table summarizing KDapp values for three independent experiments (n = 3) (right). SHOC2ΔN has impaired affinity compared to SHOC2 WT. c, Structure of SHOC2:PP1C:MRAS with non-identical residues between PP1C α,β and γ highlighted in yellow. Three non-identical residues located at complex interfaces have similar chemical properties (arrows). d, SEC traces and associated SDS-PAGE analysis showing complex formation between SHOC2, MRAS and PP1C α,β and γ isoforms. e, Left: Representative peptide dephosphorylation assay against BRAF NTpS with PP1C α,β and γ isoforms. Data points are from two time points (n = 2) (left). Right: Bar graph summarizing mean catalytic efficiency +/- SD from three independent experiments (n = 3). All isoforms show an increase in catalytic efficiency when bound to SHOC2:RAS. [PP1C] = 10 nM, [KRAS(GCP)] = 10 µM. f, Left: Representative peptide dephosphorylation assay showing SHOC2 KDapp for MRAS:PP1C α, β and γ complexes. Data points are from two time points (n = 2) (left). Right: Bar graph summarizing mean SHOC2 KDapp +/- SD from three independent experiments (n = 3). [PP1C] = 25 nM, [MRAS(GCP)] = 1 µM. g, SEC traces (top) and associated SDS-PAGE analysis (bottom) showing MRAS(GDP) can interact with PP1C α,β and γ isoforms. All SEC and SDS-PAGE data is representative of two independent experiments.
