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Forskolin and camptothecin induce a 30 kDa protein associated with melatonin production in Y79 human retinoblastoma cells

机译:福斯高林和喜树碱在Y79人视网膜母细胞瘤细胞中诱导与褪黑素产生有关的30 kDa蛋白

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摘要

The synthesis of melatonin in Xenopus retinas, chick and quail retinal cell cultures, and Y79 human retinoblastoma cells is stimulated by cAMP through a protein synthesis-dependent mechanism. In Y79 retinoblastoma cells, combined treatment with the RNA synthesis inhibitor camptothecin and agents that elevate cAMP, such as forskolin, causes a synergistic elevation of melatonin. Using two-dimensional gel analysis we have identified a 30 kDa cytosolic protein (p30) whose radiolabeling was consistently increased in parallel with increases in arylalkylamine N- acetyltransferase activity and melatonin production that were induced by forskolin and/or camptothecin. Pulse-chase experiments suggest that the elevation in radiolabeling of p30 is due to increased synthesis. Three candidate proteins found in the mammalian pineal, protein 14–3–3, malate dehydrogenase, and recoverin, do not comigrate with p30.
机译:cAMP通过蛋白质合成依赖性机制刺激非洲爪蟾视网膜,鸡和鹌鹑视网膜细胞培养物中以及Y79人视网膜母细胞瘤细胞中褪黑激素的合成。在Y79视网膜母细胞瘤细胞中,与RNA合成抑制剂喜树碱和可升高cAMP的药物(例如福司可林)联合处理可导致褪黑素协同升高。使用二维凝胶分析,我们已经确定了一个30 k​​Da的胞质蛋白(p30),其放射性标记与由佛司可林和/或喜树碱诱导的芳基烷基胺N-乙酰基转移酶活性和褪黑激素产生同时增加。脉冲追踪实验表明,p30放射性标记的升高归因于合成的增加。在哺乳动物的松果体中发现的三种候选蛋白,即蛋白14–3–3,苹果酸脱氢酶和恢复蛋白,与p30不存在竞争性。

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