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Characterisation of a cyclic peptide that binds to the RAS binding domain of phosphoinositide 3-kinase p110α.

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journal contribution
posted on 2023-02-06, 13:57 authored by Mohamed Ismail, Stephen R Martin, Roger George, Francesca Houghton, Geoff Kelly, Raphaël AG Chaleil, Panayiotis Anastasiou, Xinyue Wang, Nicola O'Reilly, Stefania Federico, Dhira Joshi, Hemavathi Nagaraj, Rachel Cooley, Ning Sze Hui, Miriam Molina-Arcas, David C Hancock, Ali Tavassoli, Julian Downward
P110α is a member of the phosphoinositide 3-kinase (PI3K) enzyme family that functions downstream of RAS. RAS proteins contribute to the activation of p110α by interacting directly with its RAS binding domain (RBD), resulting in the promotion of many cellular functions such as cell growth, proliferation and survival. Previous work from our lab has highlighted the importance of the p110α/RAS interaction in tumour initiation and growth. Here we report the discovery and characterisation of a cyclic peptide inhibitor (cyclo-CRVLIR) that interacts with the p110α-RBD and blocks its interaction with KRAS. cyclo-CRVLIR was discovered by screening a "split-intein cyclisation of peptides and proteins" (SICLOPPS) cyclic peptide library. The primary cyclic peptide hit from the screen initially showed a weak affinity for the p110α-RBD (Kd about 360 µM). However, two rounds of amino acid substitution led to cyclo-CRVLIR, with an improved affinity for p110α-RBD in the low µM (Kd 3 µM). We show that cyclo-CRVLIR binds selectively to the p110α-RBD but not to KRAS or the structurally-related RAF-RBD. Further, using biophysical, biochemical and cellular assays, we show that cyclo-CRVLIR effectively blocks the p110α/KRAS interaction in a dose dependent manner and reduces phospho-AKT levels in several oncogenic KRAS cell lines.


Crick (Grant ID: 10070, Grant title: Downward FC001070) European Research Council (Grant ID: 834692 - RASImmune, Grant title: ERC 834692 - RASImmune) Wellcome Trust (Grant ID: 103799/A/14/Z, Grant title: WT 103799/A/14/Z) Crick (Grant ID: CC1068, Grant title: STP Structural Biology) Crick (Grant ID: 10013, Grant title: STP Peptide Chemistry) Crick (Grant ID: 10316, Grant title: Bates FC001003)