| Title | The expression and purification of human rhinovirus protease 3C. | ||
| Author | Knott, J A; Orr, D C; Montgomery, D S; Sullivan, C A; Weston, A | ||
| Journal | Eur J Biochem | Publication Year/Month | 1989-Jul |
| PMID | 2546760 | PMCID | -N/A- |
| Affiliation | 1.Genetics Department, Glaxo Group Research, Greenford, Middlesex, England. | ||
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.