A BAG-1-inhibitory peptide, GO-Pep, suppresses c-Raf activity in cancer

Herein, we explore the druggability of BAG-1S (the smallest isoform) with a specific emphasis on its interaction with C-RAF. We firstly characterize the higher order structure of BAG-1S structure to determine druggable sites by HDX-MS. We then map the BAG-1S:c-Raf interface – identifying a 20 amino acid region likely to interact with c-Raf. Following which, we use mutational analysis of BAG-1S to show specific amino acids that disrupt the BAG-1S:c-Raf interaction, downstream signaling and cell growth. Finally, we develop a BAG-1S interacting peptide (from c-Raf) that disrupts BAG-1 protein:protein interactions (including C/B-RAF, CHIP and HSP70) and modified with a cell-penetrating motif which inhibits the growth of cancer cells with associated signaling inhibition and induction of apoptosis.