Figure 4
From: Functional characterisation of a novel class of in-frame insertion variants of KRAS and HRAS
Analysis of effector interaction. (A,B) Nucleotide dissociation rates (kobs) of RAS loaded with the fluorescent GTP analogue mGppNHP were determined in the presence of various concentrations of Raf-RBD and plotted against the concentration of Raf-RBD. Solid lines, data fit. (C) Affinities of the interaction as determined from the data in (A,B). (D) GTPγS loaded G-proteins together with 26 different unrelated reference baits (cb01 to cb26) were immobilised and used for precipitation of interacting proteins from HeLa cells in duplicate. The obtained precipitations were analysed by mass spectroscopy. The summed peptide intensities of identified proteins were logarithmical transformed to the basis of 10 and are represented by shades of grey. Note: intensities smaller than 104 are not obtained due to technical constrains. In total 1254 proteins were identified with more than 2 peptides. Here, only the identified known RAS effectors and a few “background” proteins are shown.
