美国华盛顿大学Dustin J. Maly等研究人员合作使用LABEL-seq进行细胞内蛋白质丰度、活性、相互作用和可成药性的多重分析。2024年10月21日，国际知名学术期刊《自然—方法学》在线发表了这一成果。

研究人员描述了一种标记带条形码并通过测序进行生化分析富集的技术（LABEL-seq），用于在人体细胞中对成组蛋白变体进行大规模平行分析。

通过利用与稳定的、编码变体的RNA条形码的RNA结合域（RBD）的细胞内自组装，LABEL-seq促进了蛋白质特性和功能的直接测量，使用简单的RBD蛋白融合的亲和富集，然后进行高通量测序以获取共同富集的条形码。

对约20000个变体效应和约1600个BRaf变体的测量显示，癌症中常见突变位点的变异对细胞内丰度的影响很小，但可以显著改变活性、蛋白质–蛋白质相互作用和可成药性。

综合分析识别出具有相似生化角色的位点网络，并能够建模变体对细胞增殖和小分子促进降解的影响。

因此，LABEL-seq能够在原生细胞环境中直接测量多个生化特性，为蛋白质功能、疾病机制和可成药性提供了新的见解。

附：英文原文

Title: Multiplexed profiling of intracellular protein abundance, activity, interactions and druggability with LABEL-seq

Author: Simon, Jessica J., Fowler, Douglas M., Maly, Dustin J.

Issue&Volume: 2024-10-21

Abstract: Here we describe labeling with barcodes and enrichment for biochemical analysis by sequencing (LABEL-seq), an assay for massively parallel profiling of pooled protein variants in human cells. By leveraging the intracellular self-assembly of an RNA-binding domain (RBD) with a stable, variant-encoding RNA barcode, LABEL-seq facilitates the direct measurement of protein properties and functions using simple affinity enrichments of RBD protein fusions, followed by high-throughput sequencing of co-enriched barcodes. Measurement of ~20,000 variant effects for ~1,600 BRaf variants revealed that variation at positions frequently mutated in cancer minimally impacted intracellular abundance but could dramatically alter activity, protein–protein interactions and druggability. Integrative analysis identified networks of positions with similar biochemical roles and enabled modeling of variant effects on cell proliferation and small molecule-promoted degradation. Thus, LABEL-seq enables direct measurement of multiple biochemical properties in a native cellular context, providing insights into protein function, disease mechanisms and druggability.

DOI: 10.1038/s41592-024-02456-7

Source: https://www.nature.com/articles/s41592-024-02456-7