2026-04-13T01:04:21Zhttps://recercat.cat/oai/request

oai:recercat.cat:10230/700032025-12-12T02:56:45Zcom_2072_6col_2072_452952

00925njm 22002777a 4500 dc Beltran, Antoni author Jiang, Xiang'er author Shen, Yue author Lehner, Ben, 1978- author 2025-03-25T07:25:42Z 2025-03-25T07:25:42Z 2025 Missense variants that change the amino acid sequences of proteins cause one-third of human genetic diseases1. Tens of millions of missense variants exist in the current human population, and the vast majority of these have unknown functional consequences. Here we present a large-scale experimental analysis of human missense variants across many different proteins. Using DNA synthesis and cellular selection experiments we quantify the effect of more than 500,000 variants on the abundance of more than 500 human protein domains. This dataset reveals that 60% of pathogenic missense variants reduce protein stability. The contribution of stability to protein fitness varies across proteins and diseases and is particularly important in recessive disorders. We combine stability measurements with protein language models to annotate functional sites across proteins. Mutational effects on stability are largely conserved in homologous domains, enabling accurate stability prediction across entire protein families using energy models. Our data demonstrate the feasibility of assaying human protein variants at scale and provides a large consistent reference dataset for clinical variant interpretation and training and benchmarking of computational methods. A.B. and B.L. were funded by a European Research Council (ERC) Advanced (883742) grant, 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, Agència de Gestió d’Ajuts Universitaris i de Recerca (AGAUR, 2017 SGR 1322), and the CERCA Program/Generalitat de Catalunya. A.B. was funded by an EMBO fellowship (ALTF 183-2020) and by the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement 101030961. X.J. and Y.S. were funded by the National Natural Science Foundation of China (32322047) and Jiangsu Provincial Department of Science and Technology (BM2023009). We thank all members of the Lehner laboratory for helpful discussions and suggestions. Clinical genetics Computational biology and bioinformatics Genomics High-throughput screening Protein folding Site-saturation mutagenesis of 500 human protein domains