西班牙巴塞罗那科技学院Ben Lehner团队近期取得重要工作进展，他们研究分析了500多个人类蛋白结构域的位点饱和诱变。相关研究成果2025年1月8日在线发表于《自然》杂志上。

据介绍，改变蛋白质氨基酸序列的错义变异会导致三分之一的人类遗传性疾病。在当前人类群体中存在着数千万种错义变异，而其中绝大多数的功能影响尚不明确。

研究人员展示了一项针对众多不同蛋白质中的人类错义变异所开展的大规模实验分析。通过DNA合成以及细胞筛选实验，研究人员量化了超过50万个变异对500多个人类蛋白结构域丰度的影响。该数据集显示，60%的致病性错义变异会降低蛋白质的稳定性。

稳定性对蛋白质适应性的贡献因蛋白质和疾病的不同而存在差异，并且在隐性遗传病中尤为重要。研究人员将稳定性测量结果与蛋白质语言模型相结合，以标注蛋白质中的功能位点。变异对稳定性的影响在同源结构域中大体上是保守的，这使得利用能量模型能够对整个蛋白质家族进行准确的稳定性预测。

总之，这一研究数据证明了大规模检测人类蛋白质变异的可行性，并为临床变异解读以及计算方法的训练和基准测试提供了一个大型且连贯一致的参考数据集。

附：英文原文

Title: Site-saturation mutagenesis of 500 human protein domains

Author: Beltran, Antoni, Jiang, Xianger, Shen, Yue, Lehner, Ben

Issue&Volume: 2025-01-08

Abstract: 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.

DOI: 10.1038/s41586-024-08370-4

Source: https://www.nature.com/articles/s41586-024-08370-4