EXPRESSION AND SELECTIVE-INHIBITION OF THE CONSTITUTIVE AND INDUCIBLE FORMS OF HUMAN CYCLOOXYGENASE

被引:372
作者
GIERSE, JK [1 ]
HAUSER, SD [1 ]
CREELY, DP [1 ]
KOBOLDT, C [1 ]
RANGWALA, SH [1 ]
ISAKSON, PC [1 ]
SEIBERT, K [1 ]
机构
[1] MONSANTO CO,SEARLE INFLAMMATORY DIS RES,ST LOUIS,MO 63167
关键词
D O I
10.1042/bj3050479
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
The enzyme cyclo-oxygenase catalyses the oxygenation of arachidonic acid, leading to the formation of prostaglandins. Recently two forms of cyclo-oxygenase have been described: a constitutive (COX-1) enzyme present in most cells and tissues, and an inducible (COX-2) isoenzyme observed in many cells in response to pro-inflammatory cytokines. Constitutive and inducible forms of human cyclo-oxygenase (hCOX-1 and hCOX-2) were cloned and expressed in insect cells, utilizing a baculovirus expression system. hCOX-1 had a specific activity of 18.8 mu mol of O-2/mg with a K-m of 13.8 mu M for arachidonate and V-max. of 1500 nmol of O-2/nmol of enzyme, whereas hCOX-2 had a specific activity of 12.2 mu mol of O-2/mg with a K-m of 8.7 mu M for arachidonate and a V-max. of 1090 nmol of O-2/nmol of enzyme. Indomethacin inhibited both hCOX-1 and hCOX-2, whereas NS-398 and Dup-697 selectively inhibited hCOX-2. Both NS-398 and Dup-697 exhibited time-dependent inactivation of hCOX-2, as did indomethacin on both enzymes. The competitive inhibitor of hCOX-1, mefenamic acid, also displayed competitive inhibition of hCOX-2. These results demonstrate the ability to generate selective non-steroidal anti-inflammatory drugs (NSAIDs), which could provide useful improvement therapeutically in the treatment of chronic inflammatory disease.
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页码:479 / 484
页数:6
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