Rostovtseva Tatiana K, Nestorovich Ekaterina M, Bezrukov Sergey M
Laboratory of Physical and Structural Biology, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892-0924 USA.
Biophys J. 2002 Jan;82(1 Pt 1):160-9. doi: 10.1016/S0006-3495(02)75383-6.
To understand the physics of polymer equilibrium and dynamics in the confines of ion channel pores, we study partitioning of poly(ethylene glycol)s (PEGs) of different molecular weights into the bacterial porin, OmpF. Thermodynamic and kinetic parameters of partitioning are deduced from the effects of polymer addition on ion currents through single OmpF channels reconstituted into planar lipid bilayer membranes. The equilibrium partition coefficient is inferred from the average reduction of channel conductance in the presence of PEG; rates of polymer exchange between the pore and the bulk are estimated from PEG-induced conductance noise. Partition coefficient as a function of polymer weight is best fitted by a "compressed exponential" with the compression factor of 1.65. This finding demonstrates that PEG partitioning into the OmpF channel pore has sharper dependence on polymer molecular weight than predictions of hard-sphere, random-flight, or scaling models. A 1360-Da polymer separates regimes of partitioning and exclusion. Comparison of its characteristic size with the size of a 2200-Da polymer previously found to separate these regimes for the alpha-toxin shows good agreement with the x-ray structural data for these channels. The PEG-induced conductance noise is compatible with the polymer mobility reduced inside the OmpF pore by an order of magnitude relatively to its value in bulk solution.
为了理解聚合物在离子通道孔内的平衡和动力学物理过程,我们研究了不同分子量的聚乙二醇(PEG)在细菌孔蛋白OmpF中的分配情况。通过将聚合物添加到重构于平面脂质双分子层膜中的单个OmpF通道上,观察其对离子电流的影响,从而推导出分配的热力学和动力学参数。平衡分配系数可从存在PEG时通道电导的平均降低值推断得出;聚合物在孔与本体之间的交换速率则根据PEG诱导的电导噪声来估计。聚合物重量与分配系数的函数关系最适合用压缩因子为1.65的“压缩指数”来拟合。这一发现表明,PEG在OmpF通道孔中的分配对聚合物分子量的依赖性比硬球、随机飞行或标度模型的预测更为明显。一种1360道尔顿的聚合物分隔了分配和排斥区域。将其特征尺寸与先前发现可分隔α-毒素这些区域的2200道尔顿聚合物的尺寸进行比较,结果与这些通道的X射线结构数据高度吻合。PEG诱导的电导噪声与OmpF孔内聚合物迁移率相对于其在本体溶液中的值降低了一个数量级相一致。