Literature DB >> 7929296

Activity of two-chain recombinant human cytomegalovirus protease.

B C Holwerda1, A J Wittwer, K L Duffin, C Smith, M V Toth, L S Carr, R C Wiegand, M L Bryant.   

Abstract

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.

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Year:  1994        PMID： 7929296

Source DB:  PubMed          Journal:  J Biol Chem        ISSN： 0021-9258            Impact factor:   5.157


  19 in total

1.  Cytomegalovirus capsid protease: biological substrates are cleaved more efficiently by full-length enzyme (pUL80a) than by the catalytic domain (assemblin).

Authors:  Steve M Fernandes; Edward J Brignole; Kanchan Taori; Wade Gibson
Journal:  J Virol       Date:  2011-01-26       Impact factor: 5.103

2.  Cleavage of human cytomegalovirus protease pUL80a at internal and cryptic sites is not essential but enhances infectivity.

Authors:  Amy N Loveland; Chee-Kai Chan; Edward J Brignole; Wade Gibson
Journal:  J Virol       Date:  2005-10       Impact factor: 5.103

3.  Enzymatic activities of human cytomegalovirus maturational protease assemblin and its precursor (pPR, pUL80a) are comparable: [corrected] maximal activity of pPR requires self-interaction through its scaffolding domain.

Authors:  Edward J Brignole; Wade Gibson
Journal:  J Virol       Date:  2007-02-07       Impact factor: 5.103

4.  Cytomegalovirus assembly protein precursor and proteinase precursor contain two nuclear localization signals that mediate their own nuclear translocation and that of the major capsid protein.

Authors:  S M Plafker; W Gibson
Journal:  J Virol       Date:  1998-10       Impact factor: 5.103

Review 5.  Protease inhibitors as antiviral agents.

Authors:  A K Patick; K E Potts
Journal:  Clin Microbiol Rev       Date:  1998-10       Impact factor: 26.132

6.  Cytomegalovirus assemblin: the amino and carboxyl domains of the proteinase form active enzyme when separately cloned and coexpressed in eukaryotic cells.

Authors:  M R Hall; W Gibson
Journal:  J Virol       Date:  1996-08       Impact factor: 5.103

7.  Independently cloned halves of cytomegalovirus assemblin, An and Ac, can restore proteolytic activity to assemblin mutants by intermolecular complementation.

Authors:  M R Hall; W Gibson
Journal:  J Virol       Date:  1997-02       Impact factor: 5.103

8.  Virus-specific interaction between the human cytomegalovirus major capsid protein and the C terminus of the assembly protein precursor.

Authors:  M Beaudet-Miller; R Zhang; J Durkin; W Gibson; A D Kwong; Z Hong
Journal:  J Virol       Date:  1996-11       Impact factor: 5.103

9.  Cytomegalovirus assemblin (pUL80a): cleavage at internal site not essential for virus growth; proteinase absent from virions.

Authors:  Chee-Kai Chan; Edward J Brignole; Wade Gibson
Journal:  J Virol       Date:  2002-09       Impact factor: 5.103

10.  Cloning, expression and characterization of the proteinase from human herpesvirus 6.

Authors:  N J Tigue; P J Matharu; N A Roberts; J S Mills; J Kay; R Jupp
Journal:  J Virol       Date:  1996-06       Impact factor: 5.103
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