Cromlish W A, Payette P, Culp S A, Ouellet M, Percival M D, Kennedy B P
Department of Biochemistry and Molecular Biology, Merck Frosst Center for Therapeutic Research, Merck Frosst Canada, Inc., Pointe-Claire-Dorval, Quebec, Canada.
Arch Biochem Biophys. 1994 Oct;314(1):193-9. 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 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.
使用重组杆状病毒在昆虫细胞中高水平表达了活性人环氧化酶-2（Cox-2）。感染细胞微粒体中Cox-2的比活性为0.51 μmol O2/分钟/毫克，与从绵羊胎盘部分纯化得到的Cox-2（0.55 μmol O2/分钟/毫克）相当。在昆虫细胞中表达的Cox-2酶有不同程度的糖基化，且大部分环氧化酶活性存在于高速微粒体沉淀中。经阿司匹林处理后，昆虫细胞表达的这种酶也表现出产生特征性15-羟基二十碳四烯酸的能力，并且对多种已知的非甾体抗炎药具有典型的抑制曲线。
