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一种高电导阳离子通道,来自自由生活的土壤细菌 Rhizobium etli 的内膜。

A high-conductance cation channel from the inner membrane of the free-living soil bacteria Rhizobium etli.

机构信息

Departamento de Biología Molecular de Plantas, Instituto de Biotecnología, UNAM, Ap. Postal 510-3, Cuernavaca, Morelos 62210, Mexico.

出版信息

Arch Microbiol. 2010 Jul;192(7):595-602. doi: 10.1007/s00203-010-0587-3. Epub 2010 May 26.

Abstract

In this communication we reported the study of a cation channel present in the cytoplasmic membrane of the nitrogen fixing bacterium Rhizobium etli. Inner-membrane (IM) vesicles were purified and fused into planar lipid bilayers (PLBs), under voltage clamp conditions. We have found that fusion of IM-enriched vesicle fractions with these model membranes leads, mainly (>30% of 46 experiments), to the reconstitution of high-conductance channels. Following this strategy, the activity of a channel with main open conductance of 198 pS, in symmetrical 100 mM KCl, was recorded. The single-channel conductance increase to 653 pS in the presence of a 5:1 (cis to trans) gradient of KCl. The channel exhibits voltage dependency and a weak selectivity for cations showing a permeability ratios of P (Rb)/P (K) = 0.96, P (Na)/P (K) = 0.07, and a conductance ratio of gamma(Rb)/gamma(K) = 1.1. The channel here characterized represents a previously undescribed Rhizobium channel although its precise role in rhizobial physiology remains yet to be determined.

摘要

在本通讯中,我们报道了一种存在于固氮菌 Rhizobium etli 细胞质膜中的阳离子通道的研究。内膜(IM)囊泡被纯化并融合到平面脂质双层(PLB)中,在电压钳条件下。我们发现,富含 IM 的囊泡部分与这些模型膜的融合主要(在 46 次实验中的>30%)导致高电导通道的重建。采用这种策略,在 100 mM KCl 的对称条件下,记录到具有 198 pS 主要开放电导的通道的活性。在存在 5:1(顺式到反式)KCl 梯度的情况下,单通道电导增加到 653 pS。该通道表现出电压依赖性和对阳离子的弱选择性,表现出 P(Rb)/P(K)=0.96、P(Na)/P(K)=0.07 和γ(Rb)/γ(K)=1.1 的电导比。这里描述的通道代表了以前未描述的 Rhizobium 通道,尽管其在根瘤菌生理学中的确切作用仍有待确定。

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