人子宫颈癌细胞系ME-180中钙/钙调蛋白依赖性一氧化氮合酶活性
Ca2+/calmodulin-dependent nitric oxide synthase activity in the human cervix carcinoma cell line ME-180.
作者信息
Werner-Felmayer G, Werner E R, Fuchs D, Hausen A, Mayer B, Reibnegger G, Weiss G, Wachter H
机构信息
Institute for Medical Chemistry and Biochemistry, University of Innsbruck, Austria.
出版信息
Biochem J. 1993 Jan 15;289 ( Pt 2)(Pt 2):357-61. doi: 10.1042/bj2890357.
Abstract
We show here that the human cervix carcinoma cell line ME-180 expresses a constitutive nitric oxide (NO) synthase, as demonstrated by formation of [3H]citrulline and nitrite. The enzyme is dependent on tetrahydrobiopterin, NADPH, flavins and Ca2+/calmodulin. Enzyme activity is located in the cytosol rather than in the membrane fraction and can be inhibited by NG-monomethyl-L-arginine (NMMA). An antiserum to NO synthase purified from porcine cerebellum inhibited the enzyme activity. ME-180 cells released NO, as was shown by stimulation of guanylate cyclase (EC 4.6.1.2) in RFL-6 detector cells; this release was stimulated 8-fold by the Ca2+ ionophore A23187 and 2-fold by increasing the intracellular tetrahydrobiopterin levels with cytokines. This is the first characterization of a Ca2+/calmodulin-dependent NO synthase activity in human epithelial-type tumour cells.
摘要
我们在此表明，人宫颈癌细胞系ME-180表达一种组成型一氧化氮（NO）合酶，这通过[3H]瓜氨酸和亚硝酸盐的形成得以证明。该酶依赖于四氢生物蝶呤、NADPH、黄素和Ca2+/钙调蛋白。酶活性位于胞质溶胶而非膜组分中，并且可被NG-单甲基-L-精氨酸（NMMA）抑制。从猪小脑纯化的针对NO合酶的抗血清可抑制该酶活性。如在RFL-6检测细胞中鸟苷酸环化酶（EC 4.6.1.2）的刺激所显示，ME-180细胞释放NO；Ca2+离子载体A23187可使这种释放增加8倍，而细胞因子增加细胞内四氢生物蝶呤水平可使其增加2倍。这是首次对人上皮型肿瘤细胞中Ca2+/钙调蛋白依赖性NO合酶活性进行的表征。
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