Department of Physics, University of Ottawa, Ottawa, Ontario, Canada.
Proteomics. 2022 Mar;22(5-6):e2100068. doi: 10.1002/pmic.202100068. Epub 2021 Dec 21.
Solid-state nanopores have been used extensively in biomolecular studies involving DNA and proteins. However, the interpretation of signals generated by the translocation of proteins or protein-DNA complexes remains challenging. Here, we investigate the behavior of monovalent streptavidin and the complex it forms with short biotinylated DNA over a range of nanopore sizes, salts, and voltages. We describe a simple geometric model that is broadly applicable and employ it to explain observed variations in conductance blockage and dwell time with experimental conditions. The general approach developed here underscores the value of nanopore-based protein analysis and represents progress toward the interpretation of complex translocation signals.
固态纳米孔在涉及 DNA 和蛋白质的生物分子研究中得到了广泛应用。然而，对于蛋白质或蛋白-DNA 复合物的迁移所产生的信号的解释仍然具有挑战性。在这里，我们研究了单价链霉亲和素及其与短链生物素化 DNA 形成的复合物在一系列纳米孔尺寸、盐和电压下的行为。我们描述了一个简单的几何模型，该模型具有广泛的适用性，并利用它来解释与实验条件相关的电导阻塞和停留时间的观察到的变化。这里所发展的一般方法强调了基于纳米孔的蛋白质分析的价值，并代表了朝着解释复杂的迁移信号的方向取得的进展。
