Bogard Andrew, Finn Pangaea W, McKinney Fulton, Flacau Ilinca M, Smith Aviana R, Whiting Rosey, Fologea Daniel
Department of Physics, Boise State University, Boise, ID 83725, USA.
Biomolecular Sciences Graduate Program, Boise State University, Boise, ID 83725, USA.
Membranes (Basel). 2021 Nov 19;11(11):897. doi: 10.3390/membranes11110897.
The electrochemical gradients established across cell membranes are paramount for the execution of biological functions. Besides ion channels, other transporters, such as exogenous pore-forming toxins, may present ionic selectivity upon reconstitution in natural and artificial lipid membranes and contribute to the electrochemical gradients. In this context, we utilized electrophysiology approaches to assess the ionic selectivity of the pore-forming toxin lysenin reconstituted in planar bilayer lipid membranes. The membrane voltages were determined from the reversal potentials recorded upon channel exposure to asymmetrical ionic conditions, and the permeability ratios were calculated from the fit with the Goldman-Hodgkin-Katz equation. Our work shows that lysenin channels are ion-selective and the determined permeability coefficients are cation and anion-species dependent. We also exploited the unique property of lysenin channels to transition to a stable sub-conducting state upon exposure to calcium ions and assessed their subsequent change in ionic selectivity. The observed loss of selectivity was implemented in an electrical model describing the dependency of reversal potentials on calcium concentration. In conclusion, our work demonstrates that this pore-forming toxin presents ionic selectivity but this is adjusted by the particular conduction state of the channels.
跨细胞膜建立的电化学梯度对于生物功能的执行至关重要。除了离子通道外,其他转运蛋白,如外源性成孔毒素,在天然和人工脂质膜中重组时可能表现出离子选择性,并有助于电化学梯度。在此背景下,我们利用电生理学方法评估了在平面双层脂质膜中重组的成孔毒素溶血素的离子选择性。通过记录通道暴露于不对称离子条件下的反转电位来确定膜电压,并根据与戈德曼-霍奇金- Katz方程的拟合计算渗透率比值。我们的工作表明,溶血素通道具有离子选择性,所确定的渗透系数取决于阳离子和阴离子种类。我们还利用了溶血素通道在暴露于钙离子时转变为稳定的亚导电状态的独特性质,并评估了其随后离子选择性的变化。在描述反转电位对钙浓度依赖性的电学模型中实现了观察到的选择性丧失。总之,我们的工作表明,这种成孔毒素具有离子选择性,但这是由通道的特定传导状态调节的。