华中科技大学李一伟团队报道了内源性生物分子凝聚物的高通量和蛋白质组范围的发现。相关研究成果于2024年3月18日发表在《自然—化学》。

哺乳动物细胞内的相分离调节与基因表达、信号传导、发育和疾病相关的生物分子凝聚体的形成。然而，大量内源性凝聚体及其候选相分离蛋白尚未以定量和高通量的方式被发现。

该文中，研究人员证明，内源性表达的生物分子凝聚体可以通过对不同寡聚状态的蛋白质进行分类，从而在细胞的蛋白质组中进行鉴定。研究人员使用体积压缩来调节细胞内蛋白质的浓度和拥挤程度，这是细胞生物分子凝聚体的物理调节因子。

蛋白质分配成凝聚体或相分离程度的变化导致蛋白质的低聚物状态的变化，这可以通过耦合密度梯度超速离心和定量质谱来检测。研究人员总共鉴定了1518种内源性凝聚蛋白，其中538种以前没有报道过。此外，研究人员证明了该策略可以识别对特定生物过程有反应的凝聚蛋白。

附：英文原文

Title: High-throughput and proteome-wide discovery of endogenous biomolecular condensates

Author: Li, Pengjie, Chen, Peng, Qi, Fukang, Shi, Jinyun, Zhu, Wenjie, Li, Jiashuo, Zhang, Peng, Xie, Han, Li, Lina, Lei, Mengcheng, Ren, Xueqing, Wang, Wenhui, Zhang, Liang, Xiang, Xufu, Zhang, Yiwei, Gao, Zhaolong, Feng, Xiaojun, Du, Wei, Liu, Xin, Xia, Limin, Liu, Bi-Feng, Li, Yiwei

Issue&Volume: 2024-03-18

Abstract: Phase separation inside mammalian cells regulates the formation of the biomolecular condensates that are related to gene expression, signalling, development and disease. However, a large population of endogenous condensates and their candidate phase-separating proteins have yet to be discovered in a quantitative and high-throughput manner. Here we demonstrate that endogenously expressed biomolecular condensates can be identified across a cell’s proteome by sorting proteins across varying oligomeric states. We employ volumetric compression to modulate the concentrations of intracellular proteins and the degree of crowdedness, which are physical regulators of cellular biomolecular condensates. The changes in degree of the partition of proteins into condensates or phase separation led to varying oligomeric states of the proteins, which can be detected by coupling density gradient ultracentrifugation and quantitative mass spectrometry. In total, we identified 1,518 endogenous condensate proteins, of which 538 have not been reported before. Furthermore, we demonstrate that our strategy can identify condensate proteins that respond to specific biological processes.

DOI: 10.1038/s41557-024-01485-1

Source: https://www.nature.com/articles/s41557-024-01485-1