Purification and partial characterization of coxsackievirus B3 2A protease expressed in Escherichia coli.
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.
Maghsoudi
Nader
N
Neuroscience Research Center, Shahid Beheshti University, M.C., Tehran, Iran. nmaghsoudi@sbmu.ac.ir
Khodagholi
Fariba
F
Sadjadi
Mahnaz
M
Zeinodini
Mehdi
M
Sabbaghian
Marjan
M
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002942:Circular Dichroism; D003001:Cloning, Molecular; D003546:Cysteine Endopeptidases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D004353:Drug Evaluation, Preclinical; D029822:Enterovirus B, Human / Q000201:enzymology* / Q000235:genetics; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D006801:Humans; D008969:Molecular Sequence Data; D011489:Protein Denaturation; D013050:Spectrometry, Fluorescence; D013696:Temperature; D014466:Ultraviolet Rays; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D015032:Zinc / Q000494:pharmacology
D004791:Enzyme Inhibitors; D014764:Viral Proteins; D003546:Cysteine Endopeptidases; C057400:picornain 2A, Picornavirus; D015032:Zinc