The expression and purification of human rhinovirus protease 3C.
Human rhinovirus type 14 protease 3C was expressed as a soluble and active protein in Escherichia coli. The protease was purified by a cationic-exchange step followed by gel filtration on a TSK 3000 column. The final yield of purified protease was in the range 0.5-1.0 mg/l culture grown to A550 = 1. 0. Sequence analysis revealed that greater than 90% of the N-terminal residues were methionine. The enzyme activity of the purified protease was measured by cleavage of a synthetic peptide representing a predicted Gln/Gly viral polyprotein cleavage site. A mutant protease (Cys146----Ser) was produced and purified in the same way. The yield of mutant protease 3C was approximately 150 micrograms/l from a culture grown to A550 = 1. 0. This mutant protease 3C did not cleave the synthetic peptide substrate.[1]References
- The expression and purification of human rhinovirus protease 3C. Knott, J.A., Orr, D.C., Montgomery, D.S., Sullivan, C.A., Weston, A. Eur. J. Biochem. (1989) [Pubmed]
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