title: High-level expression of enzymatically active bovine leukemia virus proteinase in E. coli.
abstract: An E. coli plasmid expressing efficiently an artificial precursor of bovine leukemia virus (BLV) proteinase under transcriptional control of the phage T7 promoter was constructed. The expression product accumulates in the induced E. coli cells in the form of insoluble cytoplasmic inclusions. Solubilization of the inclusions and a refolding step yield almost pure and completely self-processed proteinase. Purification to homogeneity was achieved by ion-exchange chromatography and reverse-phase HPLC. On a preparative scale, a high yield of enzymatically active proteinase was obtained. An initial study using a series of synthetic peptide substrates shows a distinct substrate specificity of BLV proteinase.
authors.affiliation: Czechoslovak Academy of Sciences, Institute of Molecular Genetics, Prague.
authors.forename: M
authors.initials: M
authors.lastname: Andreánsky
authors.forename: O
authors.initials: O
authors.lastname: Hrusková-Heidingsfeldová
authors.forename: J
authors.initials: J
authors.lastname: Sedlácek
authors.forename: J
authors.initials: J
authors.lastname: Konvalinka
authors.forename: I
authors.initials: I
authors.lastname: Bláha
authors.forename: P
authors.initials: P
authors.lastname: Jecmen
authors.forename: M
authors.initials: M
authors.lastname: Horejsí
authors.forename: P
authors.initials: P
authors.lastname: Strop
authors.forename: M
authors.initials: M
authors.lastname: Fábry
mesh_terms: D000595:Amino Acid Sequence; D010450:Endopeptidases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D015870:Gene Expression; D016333:HIV Protease / Q000378:metabolism; D015497:HIV-1 / Q000201:enzymology; D001909:Leukemia Virus, Bovine / Q000201:enzymology*; D008969:Molecular Sequence Data; D010455:Peptides / Q000378:metabolism; D010957:Plasmids; D013379:Substrate Specificity
chemical_list: D010455:Peptides; D010450:Endopeptidases; D016333:HIV Protease