Format

Send to

Choose Destination
Sci Rep. 2019 Jun 24;9(1):9150. doi: 10.1038/s41598-019-45711-0.

Simultaneous assay for protease activities of hepatitis C virus and human immunodeficiency virus based on fluorescence detection.

Author information

1
Faculty of Pharmaceutical Sciences, Nagasaki International University, 2825-7 Huis Ten Bosch machi, Sasebo, Nagasaki, 859-3298, Japan. kabashima@niu.ac.jp.
2
Department of Pathophysiology, Yokohama University of Pharmacy, 601 Matano-cho, Totsuka-ku, Yokohama-shi, Kanagawa, 245-0066, Japan.
3
Faculty of Pharmaceutical Sciences, Nagasaki International University, 2825-7 Huis Ten Bosch machi, Sasebo, Nagasaki, 859-3298, Japan.
4
Faculty of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-machi, Nagasaki, 852-8521, Japan.
5
Department of Laboratory Sciences, Gunma University Graduate School of Health Sciences, 3-39-22 Showa-machi, Maebashi-shi, Gunma, 371-8514, Japan.

Abstract

Hepatitis C virus protease (HCV-PR) and human immunodeficiency virus protease (HIV-PR) are important for virus maturation, and thus can be used as potential target molecules for the development of antiviral drugs for the treatment of viral infections. In this study, a novel assay was developed to determine HCV-PR activity. This assay is based on a fluorogenic reaction, in which peptide fragments generated from an acetyl peptide substrate by HCV-PR can be selectively converted into a fluorescent derivative, and quantified by high-performance liquid chromatography (HPLC) with fluorescent detection. Herein, several acetyl-peptides can be used as substrates for HPLC. The application of this assay was further validated by simultaneous detection of HCV-PR and HIV-PR in a reaction mixture. The proposed method can differentiate the enzyme activities of HCV-PR and HIV-PR in a sample using their corresponding substrates. The results suggest that this assay can detect various proteases by employing set of substrate peptides under the same reaction conditions.

Supplemental Content

Full text links

Icon for Nature Publishing Group Icon for PubMed Central
Loading ...
Support Center