Extended Data Fig. 2: 14-3-3 induced RAF dimer (+/- MEK) is stable in presence of ATP analog when C-terminus of RAF is phosphorylated.
From: Negative regulation of RAF kinase activity by ATP is overcome by 14-3-3-induced dimerization
a, CRAFKD(FCT) (left) and BRAFKD(CFT) co-purify as a 2:2 complex with 14-3-3 as judged by preparative SEC (top) and SDS-PAGE (bottom). b, Identification of phosphorylation on BRAF-S729 and CRAF-S621 quantified at >90% by LC-MS/MS analysis. c, Proteins co-purifying with RAFKD(FCT) identified as insect 14-3-3ζ and 14-3-3ε by LC-MS/MS analysis. d, SDS-PAGE analysis of size exclusion experiments in Fig. 2a. The corresponding SEC trace for each gel is labeled to the left, while the positions of CRAF, BRAF, MEK and 14-3-3 are labeled to the right of each gel. Elution volumes at top correspond to all gels.
