Protocol for high-throughput screening of SIRT7 inhibitors using fluorescent peptide technology
Year: 2025
Type: article
Abstract: Fluorescent peptides combine the polypeptides with fluorescent groups organically to facilitate the measurement of enzyme activity. Here, we present a protocol to evaluate Sirtuin 7 (SIRT7) enzymatic activity by analyzing changes in luminescent signals from its substrate polypeptides. We outline the steps for SIRT7 protein purification, enzymatic reaction setup, fluorescence spectrum detection, and data analysis. Additionally, we describe two applications targeting SIRT7: high-throughput screening of SIRT7 inhibitors using microplate and the validation of potential SIRT7 inhibitors. For complete details on the use and execution of this protocol, please refer to Kang et al. 1 • Large-scale purification of His-SIRT7 recombinant proteins from E. coli • Assessment of SIRT7 enzymatic activity using fluorescent peptides • High-throughput screening for identifying potential SIRT7 inhibitors • Determination of the IC 50 values for tested compounds Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Fluorescent peptides combine the polypeptides with fluorescent groups organically to facilitate the measurement of enzyme activity. Here, we present a protocol to evaluate Sirtuin 7 (SIRT7) enzymatic activity by analyzing changes in luminescent signals from its substrate polypeptides. We outline the steps for SIRT7 protein purification, enzymatic reaction setup, fluorescence spectrum detection, and data analysis. Additionally, we describe two applications targeting SIRT7: high-throughput screening of SIRT7 inhibitors using microplate and the validation of potential SIRT7 inhibitors.
Source: STAR Protocols
Institutions: Guangzhou University of Chinese Medicine, Shenzhen University
Language: English
Subfield: Geriatrics and Gerontology
Field: Medicine
Domain: Health Sciences
Open Access status: gold
Estimated APC paid: $1,550
OpenAlex ID: W4408937567