HIGH-LEVEL EXPRESSION OF ACTIVE HUMAN CYCLOOXYGENASE-2 IN INSECT CELLS

被引:43
作者
CROMLISH, WA [1 ]
PAYETTE, P [1 ]
CULP, SA [1 ]
OUELLET, M [1 ]
PERCIVAL, MD [1 ]
KENNEDY, BP [1 ]
机构
[1] MERCK FROSST CANADA INC,MERCK FROSST CTR THERAPEUT RES,DEPT BIOCHEM & MOLEC BIOL,POINTE CLAIRE H9R 4P8,PQ,CANADA
关键词
CYCLOOXYGENASE-2; OVEREXPRESSION; BACULOVIRUS; NSAIDS; GLYCOSYLATION;
D O I
10.1006/abbi.1994.1429
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 mu mol O-2/min/mg and was comparable to that obtained for partially purified Cox-2 from ovine placenta (0.55 mu mol 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. (C) 1994 Academic Press, Inc.
引用
收藏
页码:193 / 199
页数:7
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