State Key Laboratory of Bioelectronics, Southeast University, Nanjing 210096, China.
Nanoscale Res Lett. 2014 Mar 24;9(1):140. doi: 10.1186/1556-276X-9-140.
Nanopores have been proven as versatile single-molecule sensors for individual unlabeled biopolymer detection and characterization. In the present work, a relative large nanopore with a diameter of about 60 nm has been used to detect protein translocation driven by a series of applied voltages. Compared with previous studied small nanopores, a distinct profile of protein translocation through a larger nanopore has been characterized. First, a higher threshold voltage is required to drive proteins into the large nanopore. With the increase of voltages, the capture frequency of protein into the nanopore has been markedly enhanced. And the distribution of current blockage events is characterized as a function of biased voltages. Due to the large dimension of the nanopore, the adsorption and desorption phenomenon of proteins observed with a prolonged dwell time has been weakened in our work. Nevertheless, the protein can still be stretched into an unfolded state by increased electric forces at high voltages. In consideration of the high throughput of the large nanopore, a couple of proteins passing through the nanopore simultaneously occur at high voltage. As a new feature, the feasibility and specificity of a nanopore with distinct geometry have been demonstrated for sensing protein translocation, which broadly expand the application of nanopore devices.
纳米孔已被证明是一种多功能的单分子传感器,可用于检测和表征单个未标记的生物聚合物。在本工作中,使用相对较大的纳米孔(直径约 60nm)来检测一系列外加电压驱动的蛋白质转位。与之前研究的小孔相比,较大纳米孔中蛋白质转位的特征明显不同。首先,需要更高的阈值电压才能将蛋白质驱动进入大纳米孔。随着电压的增加,蛋白质进入纳米孔的捕获频率明显增强。并且电流阻断事件的分布作为偏置电压的函数进行了特征描述。由于纳米孔的大尺寸,在我们的工作中,观察到的蛋白质吸附和解吸现象随着停留时间的延长而减弱。然而,在高电压下,蛋白质仍可以通过增加的电场力被拉伸成未折叠状态。考虑到大纳米孔的高通量,在高电压下同时有几个蛋白质穿过纳米孔。作为一个新的特点,具有不同几何形状的纳米孔在感测蛋白质转位方面的可行性和特异性得到了证明,这广泛扩展了纳米孔器件的应用。