title: Expression and characterization of recombinant murine cytomegalovirus protease.
abstract: The protease domain of the murine cytomegalovirus (MCMV) M80 open reading frame was expressed in and purified from Escherichia coli. The recombinant enzyme was recovered as a mixture of active one- and two-chain forms. The two-chain enzyme was formed by internal cleavage of the one-chain enzyme at the I site. Activity measurements showed that MCMV protease cleaves R- and M-site peptide mimics with kinetics similar to those of recombinant human cytomegalovirus (HCMV) protease. Both the MCMV and HCMV proteases cleave I-site peptide substrates very poorly, but the crystal structure of the HCMV protease indicates that the cytomegalovirus I site likely resides on a solvent-exposed loop close to the active site.
authors.affiliation: Searle Discovery Research, St. Louis, Missouri 63198, USA.
authors.forename: J H
authors.initials: JH
authors.lastname: Sloan
authors.forename: J M
authors.initials: JM
authors.lastname: Loutsch
authors.forename: S Y
authors.initials: SY
authors.lastname: Boyce
authors.forename: B C
authors.initials: BC
authors.lastname: Holwerda
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D010450:Endopeptidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D015394:Molecular Structure; D018146:Muromegalovirus / Q000201:enzymology*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D011993:Recombinant Fusion Proteins; D010450:Endopeptidases