| Autor: |
Holwerda, B C, Wittwer, A J, Duffin, K L, Smith, C, Toth, M V, Carr, L S, Wiegand, R C, Bryant, M L |
| Zdroj: |
Journal of Biological Chemistry; October 1994, Vol. 269 Issue: 41 p25911-25915, 5p |
| Abstrakt: |
The human cytomegalovirus UL80 protease was expressed in Escherichia coli and purified by metal-chelate chromatography using a histidine tag engineered at the amino terminus. Cleavage of the 30-kDa protease at an internal site, VEA/A144, resulted in the recovery of 16- plus 14-kDa two-chain protease. The amino-terminal 16-kDa chain and the carboxyl-terminal 14-kDa chain remained associated as an active enzyme that was modified specifically at Ser132 on the 16-kDa chain by [3H]diisopropyl fluorophosphate. Disruption of the cleavage site by mutation from VEA/A to AEA/A facilitated the recovery of active 30-kDa one-chain enzyme that could be similarly modified at Ser132 by [3H]diisopropyl fluorophosphate. Both one- and two-chain enzymes cleaved recombinant assembly protein at the maturation site, VNA/S, and a peptide, GVVNASARL, mimicking this site. Internal processing does not inactivate the protease but forms a two-chain enzyme that retains activity. |
| Databáze: |
Supplemental Index |
| Externí odkaz: |
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