Identification of an H-Ras nanocluster disrupting peptide
: Steffen, Candy Laura; Manoharan, Ganesh Babu; Pavic, Karolina; Yeste-Vázquez, Alejandro; Knuuttila, Matias; Arora, Neha; Zhou, Young; Härmä, Harri; Gaigneaux, Anthoula; Grossmann, Tom N.; Abankwa, Daniel Kwaku
Publisher: Nature Research
: 2024
: Communications Biology
: Communications biology
: Commun Biol
: 837
: 7
: 1
: 2399-3642
: 2399-3642
DOI: https://doi.org/10.1038/s42003-024-06523-9(external)
: https://doi.org/10.1038/s42003-024-06523-9(external)
: https://research.utu.fi/converis/portal/detail/Publication/457170380(external)
Hyperactive Ras signalling is found in most cancers. Ras proteins are only active in membrane nanoclusters, which are therefore potential drug targets. We previously showed that the nanocluster scaffold galectin-1 (Gal1) enhances H-Ras nanoclustering via direct interaction with the Ras binding domain (RBD) of Raf. Here, we establish that the B-Raf preference of Gal1 emerges from the divergence of the Raf RBDs at their proposed Gal1-binding interface. We then identify the L5UR peptide, which disrupts this interaction by binding with low micromolar affinity to the B- and C-Raf-RBDs. Its 23-mer core fragment is sufficient to interfere with H-Ras nanoclustering, modulate Ras-signalling and moderately reduce cell viability. These latter two phenotypic effects may also emerge from the ability of L5UR to broadly engage with several RBD- and RA-domain containing Ras interactors. The L5UR-peptide core fragment is a starting point for the development of more specific reagents against Ras-nanoclustering and -interactors.
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The study was supported by the grant INTER/NWO/19/14061736-HRAS-PPi of the Luxembourg National Research Fund (FNR) and OCENW.KLEIN.228 of the Dutch Research Council (NWO) to DA and TG, respectively, as well as FNR-grant INTER/Mobility/2021/BM/15591725/panRAFi-PB to GM. Prof. Marc Therrien (IRIC, Université de Montréal, Canada) is gratefully acknowledged for hosting GM in his laboratory. Dr. Hugo Lavoie and Dr. Ting Jin (IRIC, Université de Montréal, Canada) are thanked for their support and advice to GM during his sabbatical. Geneviève Arseneault (IRIC, Université de Montréal, Canada) is thanked for the technical support. pET28a-L5UR was a gift from Dr. Latifa Elantak (CNRS Marseille, France). pcDNA-Hygro-Anginex was a gift from Prof. Victor L. Thijssen (VU Amsterdam, Netherlands). The KinCon plasmid was obtained from Dr. Eduard Stefan (University of Innsbruck, Austria).
