Theuvenet A P, Bindels R J
Biochim Biophys Acta. 1980 Jul;599(2):587-95. doi: 10.1016/0005-2736(80)90202-3.
A method for studying ion uptake in enzymatically isolated protoplasts from the yeast, Saccharomyces cerevisiae, is described. The kinetics of K+ and Rb+ uptake, metabolic proton extrusion and cell electrophoretic mobility have been determined. Enzymic removal of the cell wall does not significantly alter the above-mentioned properties of the yeast cells. It is concluded that studies of these properties can be performed equally well with intact yeast cells or protoplasts. However, in studies aimed at determining effects of complex organic substances, e.g., antibiotics, on plasma membrane function the use of protoplasts is recommended. The effectiveness of the antibiotic, Dio-9, for example, in reversing the metabolic proton extrusion into a net proton influx is at least 50 times higher after enzymic removal of the yeast cell wall.
本文描述了一种研究酿酒酵母酶解原生质体离子吸收的方法。测定了钾离子和铷离子吸收动力学、代谢性质子外排和细胞电泳迁移率。酶法去除细胞壁不会显著改变酵母细胞的上述特性。得出结论,对这些特性的研究使用完整酵母细胞或原生质体均可同样出色地进行。然而,在旨在确定复杂有机物质(如抗生素)对质膜功能影响的研究中,建议使用原生质体。例如,在酶法去除酵母细胞壁后,抗生素Dio-9使代谢性质子外排逆转成净质子内流的效果至少高50倍。