Identification of deubiquitinase inhibitors via high-throughput screening using a fluorogenic ubiquitin-rhodamine assay

STAR Protocols

Sara J. Buhrlage¹  Dominick Casalena²  Anthony C. Varca³  Douglas Auld⁴ 

[1] Corresponding author;Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA;Department of Cancer Biology and the Linde Program in Cancer Chemical Biology, Dana-Farber Cancer Institute, Boston, MA 02215, USA;FAST Lab, Chemical Biology and Therapeutics, Novartis Institutes for BioMedical Research, Cambridge, MA 02139, USA;

关键词: High Throughput Screening;    Molecular/Chemical Probes;    Protein Biochemistry;    Protein expression and purification;   

DOI  :  

来源: DOAJ

【 摘 要 】

Summary: 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 in vitro 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).

【 授权许可】

Unknown