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

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 develo...

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Autores principales:	Tsutomu Kabashima, Keiko Tonooka, Makoto Takada, Masaaki Kai, Takayuki Shibata
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2019
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/dcaacc38c6fd49df95cf7f9cf5f13f2f
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spelling	oai:doaj.org-article:dcaacc38c6fd49df95cf7f9cf5f13f2f2021-12-02T15:08:08ZSimultaneous assay for protease activities of hepatitis C virus and human immunodeficiency virus based on fluorescence detection10.1038/s41598-019-45711-02045-2322https://doaj.org/article/dcaacc38c6fd49df95cf7f9cf5f13f2f2019-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-019-45711-0https://doaj.org/toc/2045-2322Abstract 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.Tsutomu KabashimaKeiko TonookaMakoto TakadaMasaaki KaiTakayuki ShibataNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 9, Iss 1, Pp 1-6 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Tsutomu Kabashima Keiko Tonooka Makoto Takada Masaaki Kai Takayuki Shibata Simultaneous assay for protease activities of hepatitis C virus and human immunodeficiency virus based on fluorescence detection
description	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.
format	article
author	Tsutomu Kabashima Keiko Tonooka Makoto Takada Masaaki Kai Takayuki Shibata
author_facet	Tsutomu Kabashima Keiko Tonooka Makoto Takada Masaaki Kai Takayuki Shibata
author_sort	Tsutomu Kabashima
title	Simultaneous assay for protease activities of hepatitis C virus and human immunodeficiency virus based on fluorescence detection
title_short	Simultaneous assay for protease activities of hepatitis C virus and human immunodeficiency virus based on fluorescence detection
title_full	Simultaneous assay for protease activities of hepatitis C virus and human immunodeficiency virus based on fluorescence detection
title_fullStr	Simultaneous assay for protease activities of hepatitis C virus and human immunodeficiency virus based on fluorescence detection
title_full_unstemmed	Simultaneous assay for protease activities of hepatitis C virus and human immunodeficiency virus based on fluorescence detection
title_sort	simultaneous assay for protease activities of hepatitis c virus and human immunodeficiency virus based on fluorescence detection
publisher	Nature Portfolio
publishDate	2019
url	https://doaj.org/article/dcaacc38c6fd49df95cf7f9cf5f13f2f
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_version_	1718388286590812160

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

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