Cátedra de Biofísica, Facultad de Odontología, UBA, Buenos Aires, Argentina.
Eur Biophys J. 2011 Sep;40(9):1029-42. doi: 10.1007/s00249-011-0715-2. Epub 2011 Jun 16.
Polycystin-2 (PC2, TRPP2) is a TRP-type, non-selective cation channel whose dysfunction is implicated in changes in primary cilium structure and genesis of autosomal dominant polycystic kidney disease (ADPKD). Lithium (Li(+)) is a potent pharmaceutical agent whose effect on cell function is largely unknown. In this work, we explored the effect of Li(+) on PC2 channel function. In vitro translated PC2 was studied in a lipid bilayer reconstitution system exposed to different chemical conditions such as Li(+) or K(+) chemical gradients and different symmetrical concentrations of either cation. Li(+) inhibited PC2 function only from the external side, by decreasing the single-channel conductance and modifying the reversal potential consistent with both permeability to and blockage of the channel. When a chemical gradient was imposed, the PC2 single-channel conductance was 144 pS and 107 pS for either K(+) or Li(+), respectively. Data were analysed in terms of the Goldman-Hodgkin-Katz approximation and energy models based on absolute rate theory to understand the mechanism(s) of Li(+) transport and blockage of PC2. The 2S3B model better explained the findings, including saturation, anomalous mole fraction, non-linearity of the current-voltage curves under bi-ionic conditions and concentration dependence of permeability ratios. The data indicate that Li(+) modifies PC2 channel function, whose effect unmasks a high-affinity binding site for this ion, and an intrinsic asymmetry in the pore structure of the channel. The findings provide insights into possible mechanism(s) of Li(+) regulation of ciliary length and dysfunction mediated by this cation.
多囊蛋白-2(PC2,TRPP2)是一种 TRP 型非选择性阳离子通道,其功能障碍与初级纤毛结构的变化和常染色体显性多囊肾病(ADPKD)的发生有关。锂(Li(+))是一种有效的药物,其对细胞功能的影响尚不清楚。在这项工作中,我们探讨了 Li(+)对 PC2 通道功能的影响。在脂质双层重建系统中研究了体外翻译的 PC2,该系统暴露于不同的化学条件下,如 Li(+)或 K(+)化学梯度和两种阳离子的不同对称浓度。Li(+)仅从外部抑制 PC2 功能,通过降低单通道电导并改变反转电位,与通道的通透性和阻塞一致。当施加化学梯度时,PC2 单通道电导分别为 144 pS 和 107 pS,分别为 K(+)或 Li(+)。根据 Goldman-Hodgkin-Katz 近似和基于绝对速率理论的能量模型对数据进行分析,以了解 Li(+)转运和 PC2 阻塞的机制。2S3B 模型更好地解释了发现结果,包括饱和、异常摩尔分数、双离子条件下电流-电压曲线的非线性以及渗透率比值的浓度依赖性。数据表明,Li(+)修饰 PC2 通道功能,其作用揭示了该离子的高亲和力结合位点,以及通道孔结构的固有不对称性。这些发现为 Li(+)调节纤毛长度和由该阳离子介导的功能障碍的可能机制提供了见解。