Wu Ting, Zhou Hang-Xu, Tian Jing, Zhou Rong, Huangfu Shangwei, Jin Bi-Kun, Sablina Anna, Zhou Fangfang, Chen Hongli, Tang Shibing, Zhang Long, Yang Bing
Key Laboratory of Novel Targets and Drug Study for Neural Repair of Zhejiang Province, School of Medicine, Hangzhou City University, Hangzhou, 310015, Zhejiang, China.
Life Sciences Institute, Department of Medical Oncology, The Second Affiliated Hospital of Zhejiang University School of Medicine, Zhejiang University, Hangzhou, 310058, Zhejiang, China.
Genome Biol. 2025 Jul 15;26(1):205. doi: 10.1186/s13059-025-03669-5.
It is crucial to investigate protein functions in specific subcellular environments. Cross-linking mass spectrometry is a powerful tool to map the direct interactome of proteins by identifying inter-protein cross-links. However, it is challenging to identify inter-protein cross-linked peptides due to their low abundance.
We chemically synthesize the cross-linkers ePDES1 and ePDES2 with an alkyne group and a compound with azide linked to a phosphate group to enrich for cross-linked peptides.
Based on the high-quality cross-linking spectra of ePDES1 and ePDES2, our methods achieve the identification of hundreds of directly interacting proteins or substrates of thioredoxins in the nucleus and mitochondria.
在特定亚细胞环境中研究蛋白质功能至关重要。交联质谱法是通过识别蛋白质间交联来绘制蛋白质直接相互作用组的强大工具。然而，由于蛋白质间交联肽丰度低，鉴定它们具有挑战性。
我们化学合成了带有炔基的交联剂ePDES1和ePDES2以及一种叠氮基连接到磷酸基团的化合物，用于富集交联肽。
基于ePDES1和ePDES2的高质量交联谱，我们的方法实现了对细胞核和线粒体中数百种直接相互作用的蛋白质或硫氧还蛋白底物的鉴定。
