Characterization of SARS‐CoV main protease and identification of biologically active small molecule inhibitors using a continuous fluorescence‐based assay

Severe acute respiratory syndrome associated coronavirus main protease (SARS‐CoV M(pro)) has been proposed as a prime target for anti‐SARS drug development. We have cloned and overexpressed the SARS‐CoV M(pro) in Escherichia coli, and purified the recombinant M(pro) to homogeneity. The kinetic param...

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Autores principales: Kao, Richard Y., To, Amanda P.C., Ng, Louisa W.Y., Tsui, Wayne H.W., Lee, Terri S.W., Tsoi, Hoi-Wah, Yuen, Kwok-Yung
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2004
Materias:
Short Communications
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7134596/
https://www.ncbi.nlm.nih.gov/pubmed/15498556
http://dx.doi.org/10.1016/j.febslet.2004.09.026
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author Kao, Richard Y.
To, Amanda P.C.
Ng, Louisa W.Y.
Tsui, Wayne H.W.
Lee, Terri S.W.
Tsoi, Hoi-Wah
Yuen, Kwok-Yung
author_facet Kao, Richard Y.
To, Amanda P.C.
Ng, Louisa W.Y.
Tsui, Wayne H.W.
Lee, Terri S.W.
Tsoi, Hoi-Wah
Yuen, Kwok-Yung
author_sort Kao, Richard Y.
collection PubMed
description Severe acute respiratory syndrome associated coronavirus main protease (SARS‐CoV M(pro)) has been proposed as a prime target for anti‐SARS drug development. We have cloned and overexpressed the SARS‐CoV M(pro) in Escherichia coli, and purified the recombinant M(pro) to homogeneity. The kinetic parameters of the recombinant SARS‐CoV M(pro) were characterized by high performance liquid chromatography‐based assay and continuous fluorescence‐based assay. Two novel small molecule inhibitors of the SARS‐CoV M(pro) were identified by high‐throughput screening using an internally quenched fluorogenic substrate. The identified inhibitors have K (i) values at low μM range with comparable anti‐SARS‐CoV activity in cell‐based assays.
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spelling pubmed-71345962020-04-08 Characterization of SARS‐CoV main protease and identification of biologically active small molecule inhibitors using a continuous fluorescence‐based assay Kao, Richard Y. To, Amanda P.C. Ng, Louisa W.Y. Tsui, Wayne H.W. Lee, Terri S.W. Tsoi, Hoi-Wah Yuen, Kwok-Yung FEBS Lett Short Communications Severe acute respiratory syndrome associated coronavirus main protease (SARS‐CoV M(pro)) has been proposed as a prime target for anti‐SARS drug development. We have cloned and overexpressed the SARS‐CoV M(pro) in Escherichia coli, and purified the recombinant M(pro) to homogeneity. The kinetic parameters of the recombinant SARS‐CoV M(pro) were characterized by high performance liquid chromatography‐based assay and continuous fluorescence‐based assay. Two novel small molecule inhibitors of the SARS‐CoV M(pro) were identified by high‐throughput screening using an internally quenched fluorogenic substrate. The identified inhibitors have K (i) values at low μM range with comparable anti‐SARS‐CoV activity in cell‐based assays. John Wiley and Sons Inc. 2004-10-22 2004-09-25 /pmc/articles/PMC7134596/ /pubmed/15498556 http://dx.doi.org/10.1016/j.febslet.2004.09.026 Text en FEBS Letters 576 (2004) 1873-3468 © 2015 Federation of European Biochemical Societies This article is being made freely available through PubMed Central as part of the COVID-19 public health emergency response. It can be used for unrestricted research re-use and analysis in any form or by any means with acknowledgement of the original source, for the duration of the public health emergency.
spellingShingle Short Communications
Kao, Richard Y.
To, Amanda P.C.
Ng, Louisa W.Y.
Tsui, Wayne H.W.
Lee, Terri S.W.
Tsoi, Hoi-Wah
Yuen, Kwok-Yung
Characterization of SARS‐CoV main protease and identification of biologically active small molecule inhibitors using a continuous fluorescence‐based assay
title Characterization of SARS‐CoV main protease and identification of biologically active small molecule inhibitors using a continuous fluorescence‐based assay
title_full Characterization of SARS‐CoV main protease and identification of biologically active small molecule inhibitors using a continuous fluorescence‐based assay
title_fullStr Characterization of SARS‐CoV main protease and identification of biologically active small molecule inhibitors using a continuous fluorescence‐based assay
title_full_unstemmed Characterization of SARS‐CoV main protease and identification of biologically active small molecule inhibitors using a continuous fluorescence‐based assay
title_short Characterization of SARS‐CoV main protease and identification of biologically active small molecule inhibitors using a continuous fluorescence‐based assay
title_sort characterization of sars‐cov main protease and identification of biologically active small molecule inhibitors using a continuous fluorescence‐based assay
topic Short Communications
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7134596/
https://www.ncbi.nlm.nih.gov/pubmed/15498556
http://dx.doi.org/10.1016/j.febslet.2004.09.026
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