2026-04-17T15:00:25Zhttps://recercat.cat/oai/request

oai:recercat.cat:10230/608112025-12-12T03:00:44Zcom_2072_6col_2072_452952

00925njm 22002777a 4500 dc Weng, Chenchun author Faure, Andre J. author Escobedo, Albert author Lehner, Ben, 1978- author 2024-07-23T06:37:53Z 2024-07-23T06:37:53Z 2024 Thousands of proteins have been validated genetically as therapeutic targets for human diseases1. However, very few have been successfully targeted, and many are considered 'undruggable'. This is particularly true for proteins that function via protein-protein interactions-direct inhibition of binding interfaces is difficult and requires the identification of allosteric sites. However, most proteins have no known allosteric sites, and a comprehensive allosteric map does not exist for any protein. Here we address this shortcoming by charting multiple global atlases of inhibitory allosteric communication in KRAS. We quantified the effects of more than 26,000 mutations on the folding of KRAS and its binding to six interaction partners. Genetic interactions in double mutants enabled us to perform biophysical measurements at scale, inferring more than 22,000 causal free energy changes. These energy landscapes quantify how mutations tune the binding specificity of a signalling protein and map the inhibitory allosteric sites for an important therapeutic target. Allosteric propagation is particularly effective across the central β-sheet of KRAS, and multiple surface pockets are genetically validated as allosterically active, including a distal pocket in the C-terminal lobe of the protein. Allosteric mutations typically inhibit binding to all tested effectors, but they can also change the binding specificity, revealing the regulatory, evolutionary and therapeutic potential to tune pathway activation. Using the approach described here, it should be possible to rapidly and comprehensively identify allosteric target sites in many proteins. This work was funded by European Research Council (ERC) Advanced grant (883742), the Spanish Ministry of Science and Innovation (LCF/PR/HR21/52410004, EMBL Partnership, Severo Ochoa Centre of Excellence), the Bettencourt Schueller Foundation, the AXA Research Fund, Agencia de Gestio d’Ajuts Universitaris i de Recerca (AGAUR, 2017 SGR 1322), and the CERCA Program/Generalitat de Catalunya. C.W. was funded by an EMBO long-term fellowship (ALTF 881-2020). A.J.F. was funded by a Ramón y Cajal fellowship (RYC2021-033375-I) financed by the Spanish Ministry of Science and Innovation (MCIN/AEI/10.13039/501100011033) and the European Union (NextGenerationEU/PRTR). We thank all members of the Lehner laboratory for helpful discussions and suggestions. We thank the CRG Genomics Unit for sequencing and the Protein Technologies Unit for assistance with the SPR measurements. Biophysics Computational biology and bioinformatics Drug discovery Genomics Systems biology The energetic and allosteric landscape for KRAS inhibition