RAF1 is a crucial element of the MAPK/ERK signaling pathway. TurboID, an efficient promiscuous biotin ligase, enables the biotinylation of proteins in the vicinity, thus facilitating the study of complex cellular processes. In this study, we utilized TurboID fused to RAF1WT, RAF1K375M (kinase inactive) and RAF1CAT(catalytic domain, constitutively active) to investigate the intricate interaction network of RAF1 under various conditions.