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对酿酒酵母细胞中质膜电位变化的测量揭示了Tok1通道在膜电位维持中的重要性。

Measurements of plasma membrane potential changes in Saccharomyces cerevisiae cells reveal the importance of the Tok1 channel in membrane potential maintenance.

作者信息

Maresova Lydie, Urbankova Eva, Gaskova Dana, Sychrova Hana

机构信息

Department of Membrane Transport, Institute of Physiology, Academy of Sciences CR, Prague, Czech Republic.

出版信息

FEMS Yeast Res. 2006 Nov;6(7):1039-46. doi: 10.1111/j.1567-1364.2006.00140.x.

Abstract

K+ is one of the cations (besides protons) whose transport across the plasma membrane is believed to contribute to the maintenance of membrane potential. To ensure K+ transport, Saccharomyces cerevisiae cells possess several types of active and passive transporters mediating the K+ influx and efflux, respectively. A diS-C3(3) assay was used to compare the contributions of various potassium transporters to the membrane potential changes of S. cerevisiae cells in the exponential growth phase. Altogether, the contributions of six K+ transporters to the maintenance of a stable membrane potential were tested. As confirmed by the observed hyperpolarization of trk1 trk2 deletion strains, the diS-C3(3) assay is a suitable method for comparative studies of the membrane potential of yeast strains differing in the presence/absence of one or more cation transporters. We have shown that the presence of the Tok1 channel strongly influences membrane potential: deletion of the TOK1 gene results in significant plasma membrane depolarization, whereas strains overexpressing the TOK1 gene are hyperpolarized. We have also proved that plasma membrane potential is not the only parameter determining the hygromycin B sensitivity of yeast cells, and that the role of intracellular transporters in protecting against its toxic effects must also be considered.

摘要

钾离子(除质子外)是一种阳离子,其跨质膜的转运被认为有助于维持膜电位。为确保钾离子转运,酿酒酵母细胞拥有多种主动和被动转运蛋白,分别介导钾离子的流入和流出。采用二硫代辛二酸 - C3(3)(diS-C3(3))测定法比较了各种钾离子转运蛋白对指数生长期酿酒酵母细胞膜电位变化的贡献。总共测试了六种钾离子转运蛋白对维持稳定膜电位的贡献。正如trk1 trk2缺失菌株所观察到的超极化所证实的那样,diS-C3(3)测定法是一种适用于比较研究存在或不存在一种或多种阳离子转运蛋白的酵母菌株膜电位的方法。我们已经表明,Tok1通道的存在强烈影响膜电位:删除TOK1基因会导致质膜显著去极化,而过表达TOK1基因的菌株则会超极化。我们还证明,质膜电位不是决定酵母细胞对潮霉素B敏感性的唯一参数,还必须考虑细胞内转运蛋白在抵御其毒性作用中的作用。

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