Chen Yunyue, Wang Siyifei, Zhang Luhao, Peng Dandan, Huang Ke, Ji Baohua, Fu Junfen, Xu Yingke
Department of Biomedical Engineering, MOE Key Laboratory of Biomedical Engineering, State Key Laboratory of Extreme Photonics and Instrumentation, Zhejiang Provincial Key Laboratory of Cardio-Cerebral Vascular Detection Technology and Medicinal Effectiveness Appraisal, Zhejiang University, Hangzhou, Zhejiang, China.
Binjiang Institute of Zhejiang University, Hangzhou, Zhejiang, China.
Commun Biol. 2025 Mar 18;8(1):455. doi: 10.1038/s42003-025-07919-x.
Precise regulation of protein abundance is critical for cellular homeostasis, whose dysfunction may directly lead to human diseases. Optogenetics allows rapid and reversible control of precisely defined cellular processes, which has the potential to be utilized for regulation of protein dynamics at various scales. Here, we developed a novel optogenetics-based protein degradation system, namely Peptide-mediated OptoTrim-Away (POT) which employs expressed small peptides to effectively target endogenous and unmodified proteins. By engineering the light-induced oligomerization of the E3 ligase TRIM21, POT can rapidly trigger protein degradation via the proteasomal pathway. Our results showed that the developed POT-PI3K and POT-GPX4 modules, which used the iSH2 and FUNDC1 domains to specifically target phosphoinositide 3-kinase (PI3K) and glutathione peroxidase 4 (GPX4) respectively, were able to potently induce the degradation of these endogenous proteins by light. Both live-cell imaging and biochemical experiments validated the potency of these tools in downregulating cancer cell migration, proliferation, and even promotion of cell apoptosis. Therefore, we believe the POT offers an alternative and practical solution for rapid manipulation of endogenous protein levels, and it could potentially be employed to dissect complex signaling pathways in cell and for targeted cellular therapies.
蛋白质丰度的精确调控对于细胞稳态至关重要，其功能失调可能直接导致人类疾病。光遗传学能够对精确界定的细胞过程进行快速且可逆的控制，这有可能用于在各种尺度上调控蛋白质动态。在此，我们开发了一种基于光遗传学的新型蛋白质降解系统，即肽介导的光控去除（POT），它利用表达的小肽有效靶向内源性和未修饰的蛋白质。通过对E3连接酶TRIM21的光诱导寡聚化进行工程改造，POT能够通过蛋白酶体途径快速触发蛋白质降解。我们的结果表明，所开发的POT-PI3K和POT-GPX4模块，分别利用iSH2和FUNDC1结构域特异性靶向磷酸肌醇3激酶（PI3K）和谷胱甘肽过氧化物酶4（GPX4），能够通过光照有效诱导这些内源性蛋白质的降解。活细胞成像和生化实验均验证了这些工具在下调癌细胞迁移、增殖以及促进细胞凋亡方面的效力。因此，我们认为POT为快速操纵内源性蛋白质水平提供了一种替代且实用的解决方案，并且它有可能用于剖析细胞中的复杂信号通路以及进行靶向细胞治疗。
