Jeon Byoung-Jin, Muthukumar Murugappan
Department of Polymer Science and Engineering, University of Massachusetts, Amherst, Massachusetts 01003, United States.
Macromolecules. 2016 Dec 13;49(23):9132-9138. doi: 10.1021/acs.macromol.6b01663. Epub 2016 Nov 22.
The electrostatic origins behind the speed of translocation of a uniformly charged flexible macromolecule through -hemolysin (HL) protein pores under a voltage are investigated using variations in pH and electrolyte concentration. We have measured durations of successful threading of poly(styrenesulfonate) through HL at two different pH conditions, pH 4.5 and pH 7.5, under various salt concentration conditions. Salt concentrations in the donor () and the recipient () compartments influence the polymer translocation dynamics differently, depending on pH. At both pH 4.5 and pH 7.5, decreasing the salt concentration, , results in faster polymer translocations. On the other hand, a decrease in salt concentration, , retards the polymer transport process at pH 4.5, while at pH 7.5 the translocation time is observed to be independent of . We present a theoretical model to calculate the translocation times from the free energy of the polymer along the translocation process to describe our experimental results. We show that the charge density of the polymer inside the nanopore is significantly affected by , explaining the salt effect on the speed of polymer translocation. The salt effects are attributed to the electrostatic interaction between the polymer and the exit portion of the HL pore, which is determined by the pH of the compartment. At low pH where the net charge of the end of the HL is positive, the attractive electrostatic interaction in becomes stronger, as decreases, resulting in delays in translocation process.
利用pH值和电解质浓度的变化,研究了在电压作用下,均匀带电的柔性大分子通过α - 溶血素(HL)蛋白孔的转位速度背后的静电起源。我们测量了在不同盐浓度条件下,聚(苯乙烯磺酸盐)在两种不同pH条件(pH 4.5和pH 7.5)下成功穿过HL的持续时间。供体()和受体()隔室中的盐浓度对聚合物转位动力学的影响不同,这取决于pH值。在pH 4.5和pH 7.5时,降低盐浓度,都会导致聚合物转位更快。另一方面,降低盐浓度,在pH 4.5时会阻碍聚合物传输过程,而在pH 7.5时,转位时间与无关。我们提出了一个理论模型,根据聚合物在转位过程中的自由能来计算转位时间,以描述我们的实验结果。我们表明,纳米孔内聚合物的电荷密度受显著影响,这解释了盐对聚合物转位速度的影响。盐效应归因于聚合物与HL孔出口部分之间的静电相互作用,这由隔室的pH值决定。在低pH值下,HL末端的净电荷为正,随着降低,中的吸引性静电相互作用变得更强,导致转位过程延迟。