Protein Transport through a Narrow Solid-State Nanopore at High Voltage: Experiments and Theory

被引:126
作者
Cressiot, Benjamin
Oukhaled, Abdelghani
Patriarche, Gilles [2 ]
Pastoriza-Gallego, Manuela
Betton, Jean-Michel
Auvray, Loic [3 ]
Muthukumar, Murugappan [1 ]
Bacri, Laurent
Pelta, Juan
机构
[1] Univ Massachusetts, Dept Polymer Sci & Engn, Amherst, MA 01003 USA
[2] LPN CNRS, Marcoussis, France
[3] Univ Paris Diderot, UMR CNRS 7057, Paris, France
基金
美国国家科学基金会;
关键词
solid-state nanopore; transmission electron microscope; protein translocation; protein unfolding; Fokker-Planck model; ALPHA-HEMOLYSIN; TRANSLOCATION; MOLECULES; PEPTIDES; DENATURATION; ADSORPTION; DYNAMICS; DNA;
D O I
10.1021/nn301672g
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
We report experimentally the transport of an unfolded protein through a narrow solid-state nanopore of 3 nm diameter as a function of applied voltage. The random coil polypeptide chain is larger than the nanopore. The event frequency dependency of current blockades from 200 to 750 mV follows a van't Hoff-Arrhenius law due to the confinement of the unfolded chain. The protein is an extended conformation inside the pore at high voltage. We observe that the protein dwell time decreases exponentially at medium voltage and is inversely proportional to voltage for higher values. This is consistent with the translocation mechanism where the protein is confined in the pore, creating an entropic barrier, followed by electrophoretic transport. We compare these results to our previous work with a larger pore of 20 nm diameter. Our data suggest that electro-osmotic flow and protein adsorption on the narrowest nanopore wall are minimized. We discuss the experimental data obtained as compared with recent theory for the polyelectrolyte translocation process. This theory reproduces clearly the experimental crossover between the entropic barrier regime with medium voltage and the electrophoretic regime with higher voltage.
引用
收藏
页码:6236 / 6243
页数:8
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