Department of Cancer Biology and the Linde Program in Cancer Chemical Biology, Dana-Farber Cancer Institute, Boston, MA 02215, USA.
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA.
STAR Protoc. 2021 Oct 19;2(4):100896. doi: 10.1016/j.xpro.2021.100896. eCollection 2021 Dec 17.
Identification of selective deubiquitinase (DUB) inhibitors is critical for probe development to further understand and explore DUB biological function. Here, we detail the optimization and deployment of an fluorogenic ubiquitin-rhodamine assay to conduct high-throughput screening of a small molecule library against a panel of DUBs. In screening the compound library against multiple DUBs in parallel, we describe an approach for identifying selective DUB inhibitors and provide a roadmap for enabling selective DUB inhibitor discovery. For complete details on the use and execution of this protocol, please refer to Varca et al. (2021).
鉴定选择性去泛素化酶（DUB）抑制剂对于开发探针以进一步了解和探索 DUB 的生物学功能至关重要。在这里，我们详细介绍了荧光素泛素-罗丹明测定法的优化和应用，以针对一系列 DUB 对小分子文库进行高通量筛选。在对多种 DUB 进行平行筛选化合物库时，我们描述了一种鉴定选择性 DUB 抑制剂的方法，并为选择性 DUB 抑制剂的发现提供了路线图。有关此方案的使用和执行的完整详细信息，请参见 Varca 等人。（2021 年）。
