首页> 美国卫生研究院文献>Biochemical Journal >Presence of haemin-controlled eIF-2 alpha kinases in both undifferentiated and differentiating mouse erythroleukaemia cells.
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Presence of haemin-controlled eIF-2 alpha kinases in both undifferentiated and differentiating mouse erythroleukaemia cells.

机译:血红蛋白控制的eIF-2α激酶在未分化和分化的小鼠红白血病细胞中均存在。

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摘要

In rabbit reticulocytes, globin synthesis is regulated by a haemin-controlled translational inhibitor (HCI) which acts by phosphorylating the alpha-subunit of eukaryotic initiation factor 2 (eIF-2). With purified eIF-2 as substrate, haemin-controlled eIF-2 alpha kinases could be partially purified from cultured mouse erythroleukaemia cells (MEL cells), which can be induced in vivo to erythroid differentiation. The eIF-2 alpha kinases from both uninduced and induced MEL cells are clearly distinct from the double-stranded-RNA-activated eIF-2 alpha kinase described for many mammalian cell types. A rough quantitative estimation indicates that, on a per-cell basis, induced MEL cells contain the same amount of haemin-controlled eIF-2 alpha kinase activity as rabbit reticulocytes, whereas uninduced MEL cells contain about one-tenth as much. As to their chromatographic behavior on CM-Sephadex and DEAE-cellulose and their sensitivity towards physiological concentrations of haemin (5-10 microM), the eIF-2 alpha kinases from MEL cells are indistinguishable from HCI. They differ from HCI with respect to their response towards activating stimuli such as prolonged incubation at 37 degrees C or brief exposure to the thiol reagent N-ethylmaleimide.
机译:在兔网织红细胞中,球蛋白的合成受血红素控制的翻译抑制剂(HCI)的调节,该抑制剂通过使真核生物起始因子2(eIF-2)的α亚基磷酸化来发挥作用。以纯化的eIF-2为底物,可以从培养的小鼠红白血病细胞(MEL细胞)中部分纯化血红素控制的eIF-2α激酶,可以在体内诱导红细胞分化。来自未诱导和诱导的MEL细胞的eIF-2α激酶明显不同于针对许多哺乳动物细胞类型描述的双链RNA激活的eIF-2α激酶。粗略的定量估计表明,就每个细胞而言,诱导的MEL细胞所含的血红素控制的eIF-2α激酶活性与兔网织红细胞相同,而未诱导的MEL细胞所含的活性约为兔网织细胞的十分之一。至于它们在CM-Sephadex和DEAE-纤维素上的色谱行为以及对血红蛋白(5-10 microM)的生理浓度的敏感性,来自MEL细胞的eIF-2α激酶与HCI是无法区分的。它们与HCI的区别在于它们对激活刺激的反应,例如在37°C下长时间孵育或短暂暴露于硫醇试剂N-乙基马来酰亚胺。

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