High-level expression of active human cyclooxygenase-2 in insect cells.
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 mumol O2/min/mg and was comparable to that obtained for partially purified Cox-2 from ovine placenta (0.55 mumol O2/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-hydroxyeicosa-tetraenoic acid production after aspirin treatment and had typical inhibition profiles with a number of known nonsteroidal antiinflammatory drugs.
Cromlish
W A
WA
Department of Biochemistry and Molecular Biology, Merck Frosst Center for Therapeutic Research, Merck Frosst Canada, Inc., Pointe-Claire-Dorval, Quebec, Canada.
Payette
P
P
Culp
S A
SA
Ouellet
M
M
Percival
M D
MD
Kennedy
B P
BP
D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression; D006026:Glycoside Hydrolases / Q000494:pharmacology; D006031:Glycosylation; D006801:Humans; D006893:Hydroxyeicosatetraenoic Acids / Q000096:biosynthesis; D008861:Microsomes / Q000201:enzymology; D008969:Molecular Sequence Data; D011451:Prostaglandin-Endoperoxide Synthases / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins; D018411:Spodoptera / Q000378:metabolism*; D014162:Transfection
D000894:Anti-Inflammatory Agents, Non-Steroidal; D016861:Cyclooxygenase Inhibitors; D018076:DNA, Complementary; D006893:Hydroxyeicosatetraenoic Acids; D011994:Recombinant Proteins; C025984:15-hydroxy-5,8,11,13-eicosatetraenoic acid; D011451:Prostaglandin-Endoperoxide Synthases; D006026:Glycoside Hydrolases