High-Level Expression of Active Human Cyclooxygenase-2 in Insect Cells

• Published in: Archives of biochemistry and biophysics Vol. 314; no. 1; pp. 193 - 199
• Main Authors: Cromlish, W.A., Payette, P., Culp, S.A., Ouellet, M., Percival, M.D., Kennedy, B.P.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.10.1994
• Subjects: Animals; Anti-Inflammatory Agents, Non-Steroidal - pharmacology; Baculoviridae - genetics; Base Sequence; Cell Line; Cloning, Molecular; Cyclooxygenase Inhibitors - pharmacology; DNA, Complementary - genetics; Gene Expression; Glycoside Hydrolases - pharmacology; Glycosylation; Humans; Hydroxyeicosatetraenoic Acids - biosynthesis; Microsomes - enzymology; Molecular Sequence Data; Prostaglandin-Endoperoxide Synthases - genetics; Prostaglandin-Endoperoxide Synthases - metabolism; Recombinant Proteins; Spodoptera - metabolism; Transfection
• ISSN: 0003-9861; 1096-0384
• DOI: 10.1006/abbi.1994.1429

Active human cyclooxygenase-2 (Cox-2) was expressed at high levels in insect cells using a recombinant baculovirus. The specific activity of Cox-2 in the microsomes of infected cells was 0.51 μmol O 2/min/mg and was comparable to that obtained for partially purified Cox-2 from ovine placenta (0.55 μmol O 2/min/mg). The Cox-2 enzyme expressed in insect cells was glycosylated to varying extents and most of the cyclooxygenase activity was in the high-speed microsomal pellet. The insect-cell-expressed enzyme also showed characteristic 15-hydroxyeicosatetraenoic acid production after aspirin treatment and had typical inhibition profiles with a number of known nonsteroidal antiinflammatory drugs.