Raja Mobeen, Vales Elisabeth
Institute for Biophysics, Johannes Kepler University Linz, Altenbergerstr. 69, 4040 Linz, Austria.
Biophys Chem. 2009 Jun;142(1-3):46-54. doi: 10.1016/j.bpc.2009.03.005. Epub 2009 Mar 17.
In the current study, we found that NaCl induces fast vesicle fusion and aggregation of the bacterial potassium channel KcsA in Escherichia coli (E. coli) membranes. Conventional gel electrophoresis and tryptophan fluorescence experiments were performed to detect NaCl-induced aggregation or supramolecular complexes. Interestingly, in a planar lipid bilayer, increasing NaCl concentration dramatically increased the total internal current indicating enhanced vesicle fusion and transfer of high number of channels to the lipid bilayer. Such channels appeared in clusters of variable size which exhibited higher conductivity, increased open probability and efficient blocking by K(+)-channel blocker tetraethylammonium (TEA). Furthermore, NaCl-induced KcsA aggregation was found to be specific for E. coli membrane as compared to the artificial membrane system. Our preliminary data suggest the role of NaCl in the formation of KcsA aggregates as well as in the enhancement of membrane fusion capability in E. coli membranes.
在当前研究中,我们发现氯化钠可诱导大肠杆菌(E. coli)膜中细菌钾通道KcsA的快速囊泡融合和聚集。进行了传统的凝胶电泳和色氨酸荧光实验,以检测氯化钠诱导的聚集或超分子复合物。有趣的是,在平面脂质双分子层中,增加氯化钠浓度会显著增加总内向电流,这表明囊泡融合增强以及大量通道转移至脂质双分子层。此类通道以大小可变的簇状出现,表现出更高的电导率、增加的开放概率以及被钾通道阻滞剂四乙铵(TEA)有效阻断。此外,与人工膜系统相比,发现氯化钠诱导的KcsA聚集对大肠杆菌膜具有特异性。我们的初步数据表明氯化钠在大肠杆菌膜中KcsA聚集体的形成以及膜融合能力增强方面发挥了作用。