Abstract
Reported here is the overexpression, purification and partial characterization of recombinant coxsakievirus B3 2A protease (CVB3 2Apro) from bacterial cells transformed with a plasmid containing the CVB3 2Apro cDNA sequences. The structural investigation showed that the protein contains mostly beta-strand elements and requires Zn2+ ions as a structural component which appeared to be inhibitory if added exogenously. The purified enzyme activity was optimal at 4 degrees C and had a short half-life at physiological temperature. This feature can be the result of the presence of a high content of beta-structure and also hydrophobic residues in its structure.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Animals
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Cell Line
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Circular Dichroism
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Cloning, Molecular
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Cysteine Endopeptidases / chemistry
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Cysteine Endopeptidases / genetics
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Cysteine Endopeptidases / isolation & purification*
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Cysteine Endopeptidases / metabolism*
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Drug Evaluation, Preclinical
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Enterovirus B, Human / enzymology*
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Enterovirus B, Human / genetics
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Enzyme Inhibitors / pharmacology
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Escherichia coli / genetics*
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Gene Expression*
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Humans
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Molecular Sequence Data
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Protein Denaturation
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Spectrometry, Fluorescence
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Temperature
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Ultraviolet Rays
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Viral Proteins / chemistry
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Viral Proteins / genetics
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Viral Proteins / isolation & purification*
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Viral Proteins / metabolism*
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Zinc / pharmacology
Substances
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Enzyme Inhibitors
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Viral Proteins
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Cysteine Endopeptidases
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picornain 2A, Picornavirus
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Zinc