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首页> 外文期刊>Molecular and Cellular Biology >Osmotically induced microinjection of ricin bypasses a ricin internalization defect in a Chinese hamster ovary mutant cell line.
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Osmotically induced microinjection of ricin bypasses a ricin internalization defect in a Chinese hamster ovary mutant cell line.

机译：渗透诱导的蓖麻毒素显微注射绕过了中国仓鼠卵巢突变细胞系中的蓖麻毒素内在化缺陷。

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摘要

By osmotic lysis of pinocytic vesicles we were able to inject ricin or ricin A chain directly into the cytosol of Chinese hamster ovary cells. The lag time of 1 to 2 h before the onset of the inhibition of protein synthesis by ricin in intact cells was reduced to 15 to 30 min by this method. Preincubation of cells with a low concentration of nigericin, which was shown earlier to enhance the cytotoxicity of ricin, had no effect under this condition. Direct transfer of either intact ricin or the ricin A subunit by osmotic lysis of pinocytic vesicles into the cytosol of the ricin-resistant CHO mutant cell line 4-10 rendered the mutant 4-10 cells as sensitive to ricin as the CHO pro wild-type cells. Both the lag time and the rate of inhibition of protein synthesis in the wild-type and mutant cell lines after the introduction of ricin by osmotic lysis of pinocytic vesicles were the same. These results indicate that injection of ricin into the cytosol by osmotic lysis of pinosomes bypasses the internalization defect in the mutant cell line.
机译：通过渗透性解胞囊泡，我们能够将蓖麻毒素或蓖麻毒素A链直接注入中国仓鼠卵巢细胞的胞质中。通过这种方法，在完整细胞中蓖麻毒素开始抑制蛋白质合成之前的1到2小时的延迟时间减少到15到30分钟。在此条件下，对低浓度尼日利亚烟碱的细胞进行预培养（先前已证明可增强蓖麻毒素的细胞毒性）没有作用。完整的蓖麻毒蛋白或蓖麻毒蛋白A亚基通过渗透性囊泡渗透裂解直接转移到耐蓖麻毒的CHO突变株4-10的胞质中，使得突变株4-10细胞对ricin的敏感性与CHO pro野生型一样。细胞。通过渗透性囊泡的渗透裂解导入蓖麻毒蛋白后，野生型和突变型细胞系的滞后时间和蛋白质合成的抑制率均相同。这些结果表明，通过松质体的渗透裂解将蓖麻毒蛋白注入胞质溶胶绕过了突变细胞系的内在缺陷。

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