Vuorinen Aini, Kennedy Cassandra R, McPhie Katherine A, McCarthy William, Pettinger Jonathan, Skehel J Mark, House David, Bush Jacob T, Rittinger Katrin
Proteomics Science Technology Platform, The Francis Crick Institute, London, UK.
Molecular Structure of Cell Signalling Laboratory, The Francis Crick Institute, London, UK.
Commun Chem. 2025 Jan 15;8(1):12. doi: 10.1038/s42004-025-01410-8.
Deubiquitinating enzymes (DUBs) are key regulators of cellular homoeostasis, and their dysregulation is associated with several human diseases. The ovarian tumour protease (OTU) family of DUBs are biochemically well-characterised and of therapeutic interest, yet only a few tool compounds exist to study their cellular function and therapeutic potential. Here we present a chemoproteomics fragment screening platform for identifying novel DUB-specific hit matter, that combines activity-based protein profiling with high-throughput chemistry direct-to-biology optimisation to enable rapid elaboration of initial fragment hits against OTU DUBs. Applying these approaches, we identify an enantioselective covalent fragment for OTUD7B, and validate it using chemoproteomics and biochemical DUB activity assays.
去泛素化酶（DUBs）是细胞内稳态的关键调节因子，其失调与多种人类疾病相关。DUBs中的卵巢肿瘤蛋白酶（OTU）家族在生物化学方面已得到充分表征且具有治疗意义，但用于研究其细胞功能和治疗潜力的工具化合物却很少。在此，我们展示了一个化学蛋白质组学片段筛选平台，用于鉴定新型DUB特异性活性物质，该平台将基于活性的蛋白质谱分析与高通量化学直接到生物学优化相结合，以快速优化针对OTU DUBs的初始片段活性物质。应用这些方法，我们鉴定出一种针对OTUD7B的对映选择性共价片段，并通过化学蛋白质组学和生化DUB活性测定对其进行了验证。
