title: Purification and characterization of cysteine proteinase from a baculovirus gene.
abstract: To analyze the degradation of product proteins at the late stage of virus infection in the baculovirus expression system, a cysteine proteinase was purified from hemolymph of Bombyx mori infected with wild-type B. mori nuclear polyhedorosis virus (BmNPV). The purified cysteine proteinase preparation had two protein bands (major 35-kDa active protein and 28-kDa inactive protein) on SDS-PAGE. Based on the N-terminal amino acid sequences of them, it was found that both proteins originated in the cysteine proteinase gene of BmNPV. The purified cysteine proteinase had an optimum pH at 4.0, and also had activities at neutral pHs. When recombinant luciferase was used as a natural substrate, it was degraded rapidly by the cysteine proteinase at the physiological pH of hemolymph. These results suggest that the cysteine proteinase from a BmNPV gene participates in the degradation of foreign protein expressed by the baculovirus system.
authors.affiliation: Department of Applied Biology, Faculty of Textile Science, Kyoto Institute of Technology, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Takahashi
authors.forename: S
authors.initials: S
authors.lastname: Ushiyama
authors.forename: T
authors.initials: T
authors.lastname: Suzuki
authors.forename: K
authors.initials: K
authors.lastname: Ogawa
authors.forename: K
authors.initials: K
authors.lastname: Oda
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001329:Autolysis; D016367:Baculoviridae / Q000201:enzymology* / Q000235:genetics*; D012831:Bombyx / Q000201:enzymology*; D003546:Cysteine Endopeptidases / Q000032:analysis / Q000235:genetics* / Q000302:isolation & purification*; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005814:Genes, Viral / Q000235:genetics*; D006454:Hemoglobins / Q000378:metabolism; D006458:Hemolymph / Q000201:enzymology; D006863:Hydrogen-Ion Concentration; D008156:Luciferases / Q000378:metabolism; D008969:Molecular Sequence Data; D013379:Substrate Specificity; D014764:Viral Proteins / Q000032:analysis / Q000302:isolation & purification
chemical_list: D015853:Cysteine Proteinase Inhibitors; D006454:Hemoglobins; D014764:Viral Proteins; D008156:Luciferases; D003546:Cysteine Endopeptidases