| Autor: |
Knott, Jacqueline A., Orr, David C., Montgomery, Douglas S., Sullivan, Carole A., Weston, Anthony |
| Předmět: |
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| Zdroj: |
European Journal of Biochemistry; 7/1/89, Vol. 182 Issue 3, p547-555, 9p |
| Abstrakt: |
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 µg/l from a culture grown to A550 = 1.0. This mutant protease 3C did not cleave the synthetic peptide substrate.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 µg/l from a culture grown to A550 = 1.0. This mutant protease 3C did not cleave the synthetic peptide substrate. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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