Lever J E, Seegmiller J E
J Cell Physiol. 1976 Jul;88(3):343-52. doi: 10.1002/jcp.1040880310.
Diploid human lymphoblastoid cells with altered response to ouabain inhibition of the (Na+ + K+)-dependent ATPase transport system, manifest both in whole cells and in purified plasma membrane vesicles, were selected for their resistance to 0.1 muM ouabain. Ouabain-resistant (OUA(R)) cells with normal growth at 50 times this dose were recovered at a frequency 1 X 10(-6). This frequency was increased 9-fold after exposure to ethyl methane sulphonate but was decreased by the frameshift mutagen ICR-191, under conditions where both increased the frequency of 8-azaguanine-resistant colonies. The ouabain resistance phenotype was stable after 200 population doublings in the absence of ouabain. OUA(R) clones show showed 30-50% of the wild type amount of 3H-ouabain bound per cell, with the same dissociation constant for ouabain, 0.1 muM at 0.5 mM K+, as observed in wild-type cells. Both the initial rate of uptake of 86Rb+ in OUA(R) cells and the (Na+ + K+)-dependent ATPase activity of OUA(R) plasma membranes showed decreased sensitivity to ouabain inhibition. However, growth and transport properties of OUA(R) cells in the absence of ouabain were unchanged compared with wild type cells.
对哇巴因抑制(Na⁺ + K⁺)依赖性ATP酶转运系统反应改变的二倍体人淋巴母细胞,在全细胞和纯化的质膜囊泡中均有表现,因其对0.1 μM哇巴因具有抗性而被挑选出来。在50倍此剂量下能正常生长的耐哇巴因(OUA(R))细胞以1×10⁻⁶的频率被回收。在暴露于甲磺酸乙酯后,该频率增加了9倍,但在移码诱变剂ICR - 191作用下频率降低,而在这两种情况下8 - 氮杂鸟嘌呤抗性菌落的频率均增加。在无哇巴因的情况下经过200次群体倍增后,哇巴因抗性表型是稳定的。每个OUA(R)克隆细胞显示结合的³H - 哇巴因量为野生型的30 - 50%,与野生型细胞一样,在0.5 mM K⁺时哇巴因的解离常数为0.1 μM。OUA(R)细胞中⁸⁶Rb⁺的初始摄取速率以及OUA(R)质膜的(Na⁺ + K⁺)依赖性ATP酶活性对哇巴因抑制的敏感性均降低。然而,与野生型细胞相比,在无哇巴因的情况下OUA(R)细胞的生长和转运特性未发生变化。