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Characterization of SARS‐CoV main protease and identification of biologically active small molecule inhibitors using a continuous fluorescence‐based assay
Author(s) -
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
Publication year - 2004
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2004.09.026
Subject(s) - protease , recombinant dna , escherichia coli , chemistry , covid-19 , chromatography , coronavirus , enzyme , biochemistry , biology , medicine , gene , disease , pathology , infectious disease (medical specialty)
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.