In vivo and in vitro expression of a non-mammalian cyclooxygenase-1

David W Reed , William S Bradshaw , Weilin Xie , Daniel L Simmons ∗
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引用次数: 20

Abstract

Unlike cyclooxygenase 2 (COX-2), COX-1 has never been identified, purified or cloned in a non-mammalian species. Here we report the RT-PCR cloning of a chicken cDNA that encodes the amphipathic membrane binding region and parts of the dimerization and catalytic domains of COX1-like enzyme. Sequence comparison showed this putative COX-1 to be evolutionarily less conserved than COX-2. Furthermore, whereas COX-1 in mammals is broadly expressed in tissues as a constitutive enzyme, the mRNA detected by our clone in chicken was almost absent in tissues and embryo fibroblasts (CEF). Highest expression was in brain and seminal vesicle. This transcript was not detectable during chick embryogenesis and, as is the case for mammalian COX-1, was not induced above background by mitogen stimulation. The identification of an avian COX-1 shows that COX-1 and COX-2 existed as separate catalysts for prostaglandin synthesis before the divergence of birds and mammals.

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非哺乳动物环氧合酶-1的体内外表达
与环氧合酶2 (COX-2)不同,COX-1从未在非哺乳动物物种中被鉴定、纯化或克隆。在这里,我们报道了RT-PCR克隆的鸡cDNA编码的两亲膜结合区和部分二聚化和催化结构域的cox样酶。序列比较表明,这一假定的COX-1在进化上的保守性低于COX-2。此外,COX-1在哺乳动物中作为一种组成酶在组织中广泛表达,而在鸡克隆中检测到的mRNA在组织和胚胎成纤维细胞(CEF)中几乎不存在。在脑和精囊中表达量最高。该转录本在鸡胚发育过程中未被检测到,并且与哺乳动物COX-1的情况一样,在有丝分裂原刺激的背景下未被诱导。鸟类COX-1的鉴定表明,在鸟类和哺乳动物分化之前,COX-1和COX-2分别作为前列腺素合成的催化剂存在。
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