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Enzymatic activity of the SARS coronavirus main proteinase dimer
Author(s) -
Graziano Vito,
McGrath William J.,
DeGruccio Ann Marie,
Dunn John J.,
Mangel Walter F.
Publication year - 2006
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.2006.04.004
Subject(s) - enzyme , coronavirus , dimer , biochemistry , proteinase 3 , cleavage (geology) , chemistry , escherichia coli , enzyme assay , microbiology and biotechnology , biology , covid-19 , gene , myeloperoxidase , medicine , paleontology , disease , pathology , fracture (geology) , infectious disease (medical specialty) , immunology , inflammation , organic chemistry
The enzymatic activity of the SARS coronavirus main proteinase dimer was characterized by a sensitive, quantitative assay. The new, fluorogenic substrate, (Ala‐Arg‐Leu‐Gln‐NH) 2 ‐Rhodamine, contained a severe acute respiratory syndrome coronavirus (SARS CoV) main proteinase consensus cleavage sequence and Rhodamine 110, one of the most detectable compounds known, as the reporter group. The gene for the enzyme was cloned in the absence of purification tags, expressed in Escherichia coli and the enzyme purified. Enzyme activity from the SARS CoV main proteinase dimer could readily be detected at low pM concentrations. The enzyme exhibited a high K m , and is unusually sensitive to ionic strength and reducing agents.