Biomaterial Database

High-throughput and proteome-wide discovery of endogenous biomolecular condensates. 2024

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

The Key Laboratory for Biomedical Photonics of MOE at Wuhan National Laboratory for Optoelectronics - Hubei Bioinformatics and Molecular Imaging Key Laboratory, Department of Biomedical Engineering, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, Hubei Province, China.

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.