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Solid-State Nanopore Distinguishes Ferritin and Apo-Ferritin with Identical Exteriors through Amplified Flexibility at Single-Molecule Level
Analytical Chemistry ( IF 6.7 ) Pub Date : 2023-11-02 , DOI: 10.1021/acs.analchem.3c02041 Yun-Dong Yin 1 , Fang-Fang Chen 1 , Jun Hu 1 , Lei Yang 1 , Xi-Tong Song 1 , Guo-Rong Wu 1 , Ming Xu 1 , Zhi-Yuan Gu 1
Analytical Chemistry ( IF 6.7 ) Pub Date : 2023-11-02 , DOI: 10.1021/acs.analchem.3c02041 Yun-Dong Yin 1 , Fang-Fang Chen 1 , Jun Hu 1 , Lei Yang 1 , Xi-Tong Song 1 , Guo-Rong Wu 1 , Ming Xu 1 , Zhi-Yuan Gu 1
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Protein identification and discrimination at the single-molecule level are big challenges. Solid-state nanopores as a sensitive biosensor have been used for protein analysis, although it is difficult to discriminate proteins with similar structures in the traditional discrimination method based on the current blockage fraction. Here, we select ferritin and apo-ferritin as the model proteins that exhibit identical exterior and different interior structures and verify the practicability of their discrimination with flexibility features by the strategy of gradually decreasing the nanopore size. We show that the larger nanopore (relative to the protein size) has no obvious effect on discriminating two proteins. Then, the comparable-sized nanopore plays a key role in discriminating two proteins based on the dwell time and fraction distribution, and the conformational changes of both proteins are also studied with this nanopore. Finally, in the smaller nanopore, the protein molecules are trapped rather than translocated, where two proteins are obviously discriminated through the current fluctuation caused by the vibration of proteins. This strategy has potential in the discrimination of other important similar proteins.
中文翻译：
固态纳米孔通过在单分子水平上增强灵活性来区分具有相同外观的铁蛋白和脱铁铁蛋白
单分子水平上的蛋白质识别和区分是巨大的挑战。固态纳米孔作为一种灵敏的生物传感器已被用于蛋白质分析，尽管基于当前阻塞分数的传统区分方法很难区分具有相似结构的蛋白质。在这里，我们选择铁蛋白和脱铁铁蛋白作为具有相同外部结构和不同内部结构的模型蛋白，并通过逐渐减小纳米孔尺寸的策略验证其具有柔性特征的区分的实用性。我们表明，较大的纳米孔（相对于蛋白质大小）对区分两种蛋白质没有明显影响。然后，大小相当的纳米孔在根据停留时间和分数分布区分两种蛋白质方面发挥着关键作用，并且还用该纳米孔研究了两种蛋白质的构象变化。最后，在较小的纳米孔中，蛋白质分子被捕获而不是移位，其中两种蛋白质通过蛋白质振动引起的电流波动明显区分。该策略在区分其他重要的相似蛋白质方面具有潜力。
更新日期：2023-11-02
中文翻译：
固态纳米孔通过在单分子水平上增强灵活性来区分具有相同外观的铁蛋白和脱铁铁蛋白
单分子水平上的蛋白质识别和区分是巨大的挑战。固态纳米孔作为一种灵敏的生物传感器已被用于蛋白质分析，尽管基于当前阻塞分数的传统区分方法很难区分具有相似结构的蛋白质。在这里，我们选择铁蛋白和脱铁铁蛋白作为具有相同外部结构和不同内部结构的模型蛋白，并通过逐渐减小纳米孔尺寸的策略验证其具有柔性特征的区分的实用性。我们表明，较大的纳米孔（相对于蛋白质大小）对区分两种蛋白质没有明显影响。然后，大小相当的纳米孔在根据停留时间和分数分布区分两种蛋白质方面发挥着关键作用，并且还用该纳米孔研究了两种蛋白质的构象变化。最后，在较小的纳米孔中，蛋白质分子被捕获而不是移位，其中两种蛋白质通过蛋白质振动引起的电流波动明显区分。该策略在区分其他重要的相似蛋白质方面具有潜力。
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