title: Expression of prostaglandin endoperoxide synthase-1 in a baculovirus system.
abstract: A cDNA coding for ovine prostaglandin endoperoxide (PGH) synthase-1 was used to construct a recombinant baculovirus which was expressed in Spodoptera frugiperda (Sf9) insect cells. Two proteins reactive with anti-PGH synthase antibody were produced. A larger protein (Mr = 72,000) coelectrophoresed with native enzyme; a smaller, more abundant protein (Mr = 66,000) was unglycosylated enzyme. About 90% of both the immunoreactivity and the cyclooxygenase activity were present in a low speed (10(5) g x min) pellet; variable but low peroxidase activities were observed in this fraction. The specific cyclooxygenase activity of solubilized PGH synthase-1 from Sf9 cells was 56 units/mg versus 112 units/mg for the same cDNA expressed in cos-1 cells. The baculovirus-insect cell system is not ideal for generating large amounts of active PGH synthase-1 apparently because of inefficient N-glycosylation.
authors.affiliation: Department of Biochemistry, Michigan State University, East Lansing 48824.
authors.forename: T
authors.initials: T
authors.lastname: Shimokawa
authors.forename: W L
authors.initials: WL
authors.lastname: Smith
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002478:Cells, Cultured; D015870:Gene Expression; D005822:Genetic Vectors; D011451:Prostaglandin-Endoperoxide Synthases / Q000235:genetics*; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000235:genetics; D012756:Sheep
chemical_list: D011994:Recombinant Proteins; D011451:Prostaglandin-Endoperoxide Synthases