Jisaka M, Kim R B, Boeglin W E, Nanney L B, Brash A R
Division of Clinical Pharmacology, Department of Pharmacology, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-6602, USA.
J Biol Chem. 1997 Sep 26;272(39):24410-6. doi: 10.1074/jbc.272.39.24410.
One of the effects of topical application of phorbol ester to mouse skin is the induction of an 8S-lipoxygenase in association with the inflammatory response. Here we report the molecular cloning and characterization of this enzyme. The cDNA was isolated by polymerase chain reaction from mouse epidermis and subsequently from a mouse epidermal cDNA library. The cDNA encodes a protein of 677 amino acids with a calculated molecular mass of 76 kDa. The amino acid sequence has 78% identity to a 15S-lipoxygenase cloned recently from human skin and approximately 40% identity to other mammalian lipoxygenases. When expressed in vaccinia virus-infected Hela cells, the mouse enzyme converts arachidonic acid exclusively to 8S-hydroperoxyeicosatetraenoic acid while linoleic acid is converted to 9S-hydroperoxy-linoleic acid in lower efficiency. Phorbol ester treatment of mouse skin is associated with strong induction of 8S-lipoxygenase mRNA and protein. By Northern analysis, expression of 8S-lipoxygenase mRNA was also detected in brain. Immunohistochemical analysis of phorbol ester-treated mouse skin showed the strongest reaction to 8S-lipoxygenase in the differentiated epidermal layer, the stratum granulosum. The inducibility may be a characteristic feature of the mouse 8S-lipoxygenase and its human 15S-lipoxygenase homologue.
将佛波酯局部涂抹于小鼠皮肤的作用之一是在炎症反应过程中诱导产生一种8S-脂氧合酶。在此我们报道这种酶的分子克隆及特性。通过聚合酶链反应从小鼠表皮随后又从小鼠表皮cDNA文库中分离出了cDNA。该cDNA编码一种含677个氨基酸的蛋白质，计算分子量为76 kDa。其氨基酸序列与最近从人皮肤克隆出的15S-脂氧合酶有78%的同源性，与其他哺乳动物脂氧合酶约有40%的同源性。当在痘苗病毒感染的Hela细胞中表达时，小鼠酶将花生四烯酸仅转化为8S-氢过氧化二十碳四烯酸，而将亚油酸以较低效率转化为9S-氢过氧化亚油酸。佛波酯处理小鼠皮肤与8S-脂氧合酶mRNA和蛋白质的强烈诱导相关。通过Northern分析，在脑中也检测到了8S-脂氧合酶mRNA的表达。对佛波酯处理的小鼠皮肤进行免疫组织化学分析显示，在分化的表皮层即颗粒层中对8S-脂氧合酶的反应最强。这种诱导性可能是小鼠8S-脂氧合酶及其人15S-脂氧合酶同源物的一个特征。
