An ultrasensitive system for measuring the USPs and OTULIN activity using Nanoluc as a reporter

作者：Chen, Yunfei; Wang, Lufan; Cheng, Xiaomu; Ge, Xin; Wang, Ping^*

来源：Biochemical and Biophysical Research Communications, 2014, 455(3-4): 178-183.

DOI：10.1016/j.bbrc.2014.10.139

• Ubiquitin
• Deubiquitinating enzymes
• Nanoluc
• PR619
• OTULIN

The deubiquitinating enzymes (DUBS) are a family of isopeptidases responsible for removing the ubiquitin from the ubiquitinated proteins. Identification of inhibitors for DUBs is emerging as an efficient way for discovering potential medicines for disease treatment. However, the high throughput screening (HTS) assay is still not available for all USPs, especially OTULIN. Here, we described a novel steadily quantifiable DUBS assay platform using Nanoluc (Nluc) as reporter. We further demonstrated that the Ub-Nluc assay could be used for HTS of DUBs inhibitors. Moreover, we generated a sensitive system for OTULIN inhibitors screening using Nluc as a reporter. In summary, our data indicate that Ub-Nluc and the improved Ub-Ub-GS-Nluc assay are efficient systems for measuring activities and screening inhibitors of USPs and OTULIN.

• 出版日期2014-12-12
• 单位华东师范大学