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膜电位变化作为聚-L-赖氨酸处理导致质膜形成选择性孔道的一种证明

Changes in Membrane Potential as a Demonstration of Selective Pore Formation in the Plasmalemma by Poly-l-Lysine Treatment.

作者信息

Reuveni M, Lerner H R, Poljakoff-Mayber A

机构信息

Department of Botany, Institute of Life Sciences, The Hebrew University of Jerusalem, 91904, Jerusalem, Israel.

出版信息

Plant Physiol. 1985 Oct;79(2):406-10. doi: 10.1104/pp.79.2.406.

Abstract

A technique which allows determination of solute pool concentrations in the cytosol was developed exploiting the interaction between a polycation and the anionic sites of the plasmalemma. It was shown that treatment of Nicotiana tabacum, cv Xanthi, cells in suspension culture with an appropriate concentration of poly-l-lysine induced pore formation selectively in the plasmalemma. The data presented in this paper shows that the plasmalemma of all the cells was affected while the tonoplast remained undamaged. This conclusion is based on the facts that treatment of the cells with the minimum amount of poly-l-lysine which just abolishes the electrogenic potential (similarly to carbonyl cyanide-p-trifluormethoxyphenylhydrazone and NaN(3)) induces the leakage of only a small fraction of the K(+) present in the cells. These effects of poly-l-lysine differ from the effects of polymyxin B which induces total leakage of low molecular weight solutes (R. Weimberg, H. R. Lerner, A. Poljakoff-Mayber 1983 J Exp Bot 34: 1333-1346) and therefore affects also the tonoplast.Membrane potential was determined using the partition of the lipophilic cation tetraphenylphosphonium. The electrogenic component of the membrane potential was estimated using carbonyl cyanide-p-trifluormethoxyphenylhydrazone and azide. Poly-l-lysine treatment was used to measure K(+) compartmentation in Nicotiana cells grown in a NaCl-containing medium.

摘要

利用聚阳离子与质膜阴离子位点之间的相互作用,开发了一种可测定细胞质溶质池浓度的技术。结果表明,用适当浓度的聚-L-赖氨酸处理悬浮培养的烟草品种黄花烟草细胞,可在质膜上选择性地诱导形成孔。本文提供的数据表明,所有细胞的质膜均受到影响,而液泡膜未受损。这一结论基于以下事实:用刚好消除生电电位的最小量聚-L-赖氨酸处理细胞(类似于羰基氰化物-对三氟甲氧基苯腙和NaN₃),仅诱导细胞中一小部分K⁺泄漏。聚-L-赖氨酸的这些作用不同于多粘菌素B的作用,多粘菌素B会诱导低分子量溶质完全泄漏(R. 魏姆贝格、H. R. 勒纳、A. 波利亚科夫-梅伯 1983年 《实验植物学杂志》34卷:1333 - 1346页),因此也会影响液泡膜。使用亲脂性阳离子四苯基鏻的分配来测定膜电位。使用羰基氰化物-对三氟甲氧基苯腙和叠氮化物来估计膜电位的生电成分。聚-L-赖氨酸处理用于测量在含NaCl培养基中生长的烟草细胞中的K⁺区室化。

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