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Expression and biochemical characterization of nsP2 cysteine protease of Chikungunya virus

Chikungunya virus (CHIKV) is a mosquito-borne alphavirus that causes epidemic fever, rash and polyarthralgia in Africa and Asia. Although it is known since the 1950s, new epidemiological and clinical features reported during the recent outbreak in the Indian Ocean can be regarded as the emergence of...

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Autores principales: Pastorino, Boris A.M., Peyrefitte, Christophe N., Almeras, Lionel, Grandadam, Marc, Rolland, Dominique, Tolou, Hugues J., Bessaud, Maël
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier B.V. 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7114110/
https://www.ncbi.nlm.nih.gov/pubmed/17961784
http://dx.doi.org/10.1016/j.virusres.2007.09.009
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author Pastorino, Boris A.M.
Peyrefitte, Christophe N.
Almeras, Lionel
Grandadam, Marc
Rolland, Dominique
Tolou, Hugues J.
Bessaud, Maël
author_facet Pastorino, Boris A.M.
Peyrefitte, Christophe N.
Almeras, Lionel
Grandadam, Marc
Rolland, Dominique
Tolou, Hugues J.
Bessaud, Maël
author_sort Pastorino, Boris A.M.
collection PubMed
description Chikungunya virus (CHIKV) is a mosquito-borne alphavirus that causes epidemic fever, rash and polyarthralgia in Africa and Asia. Although it is known since the 1950s, new epidemiological and clinical features reported during the recent outbreak in the Indian Ocean can be regarded as the emergence of a new disease. Numerous severe forms of the infection have been described that put emphasis on the lack of efficient antiviral therapy. Among the virus-encoded enzymes, nsP2 constitutes an attractive target for the development of antiviral drugs. It is a multifunctional protein of approximately 90 kDa with a helicase motif in the N-terminal portion of the protein while the papain-like protease activity resides in the C-terminal portion. The nsP2 proteinase is an essential enzyme whose proteolytic activity is critical for virus replication. In this work, a recombinant CHIKV nsP2pro and a C-terminally truncated variant were expressed in Escherichia coli and purified by metal–chelate chromatography. The enzymatic properties of the proteinase were then determined using specific synthetic fluorogenic substrates. This study constitutes the first characterization of a recombinant CHIKV nsP2 cysteine protease, which may be useful for future drug screening.
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spelling pubmed-71141102020-04-02 Expression and biochemical characterization of nsP2 cysteine protease of Chikungunya virus Pastorino, Boris A.M. Peyrefitte, Christophe N. Almeras, Lionel Grandadam, Marc Rolland, Dominique Tolou, Hugues J. Bessaud, Maël Virus Res Article Chikungunya virus (CHIKV) is a mosquito-borne alphavirus that causes epidemic fever, rash and polyarthralgia in Africa and Asia. Although it is known since the 1950s, new epidemiological and clinical features reported during the recent outbreak in the Indian Ocean can be regarded as the emergence of a new disease. Numerous severe forms of the infection have been described that put emphasis on the lack of efficient antiviral therapy. Among the virus-encoded enzymes, nsP2 constitutes an attractive target for the development of antiviral drugs. It is a multifunctional protein of approximately 90 kDa with a helicase motif in the N-terminal portion of the protein while the papain-like protease activity resides in the C-terminal portion. The nsP2 proteinase is an essential enzyme whose proteolytic activity is critical for virus replication. In this work, a recombinant CHIKV nsP2pro and a C-terminally truncated variant were expressed in Escherichia coli and purified by metal–chelate chromatography. The enzymatic properties of the proteinase were then determined using specific synthetic fluorogenic substrates. This study constitutes the first characterization of a recombinant CHIKV nsP2 cysteine protease, which may be useful for future drug screening. Elsevier B.V. 2008-02 2007-10-25 /pmc/articles/PMC7114110/ /pubmed/17961784 http://dx.doi.org/10.1016/j.virusres.2007.09.009 Text en Copyright © 2007 Elsevier B.V. All rights reserved. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.
spellingShingle Article
Pastorino, Boris A.M.
Peyrefitte, Christophe N.
Almeras, Lionel
Grandadam, Marc
Rolland, Dominique
Tolou, Hugues J.
Bessaud, Maël
Expression and biochemical characterization of nsP2 cysteine protease of Chikungunya virus
title Expression and biochemical characterization of nsP2 cysteine protease of Chikungunya virus
title_full Expression and biochemical characterization of nsP2 cysteine protease of Chikungunya virus
title_fullStr Expression and biochemical characterization of nsP2 cysteine protease of Chikungunya virus
title_full_unstemmed Expression and biochemical characterization of nsP2 cysteine protease of Chikungunya virus
title_short Expression and biochemical characterization of nsP2 cysteine protease of Chikungunya virus
title_sort expression and biochemical characterization of nsp2 cysteine protease of chikungunya virus
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7114110/
https://www.ncbi.nlm.nih.gov/pubmed/17961784
http://dx.doi.org/10.1016/j.virusres.2007.09.009
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