Use of a solid‐state nanopore for profiling the transferrin receptor protein and distinguishing between transferrin receptor and its ligand protein
Use of a solid‐state nanopore for profiling the transferrin receptor protein and distinguishing between transferrin receptor and its ligand protein
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使用固态纳米孔分析转铁蛋白受体蛋白并区分转铁蛋白受体及其配体蛋白
DOI：
10.1002/elps.202200147
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发表时间：
2022
期刊：
影响因子：
2.9
通讯作者：
Kim, Min Jun
中科院分区：
文献类型：
--
作者：
O'Donohue, Matthew;Saharia, Jugal;Bandara, Nuwan;Alexandrakis, Georgios;Kim, Min Jun
当被分析物在外加电场的影响下通过传感孔（纳米尺寸的通孔）移位时，纳米孔装置能够提供单分子水平的信息。在这项研究中，氮化硅（SixNy）纳米孔被用来表征不同施加电压下的人血清转铁蛋白受体蛋白（TfR）。发现随着外加电场的增加，二聚体形式的TfR呈指数减少。对单体TfR的进一步分析还表明，其展开行为正依赖于施加的电压。此外，对TfR单体及其配体蛋白——人血清转铁蛋白（hSTf）的数据进行比较发现，尽管这两种蛋白的分子量几乎相同，但可以通过固态纳米孔（SSN）来区分。最后，在每个电压下实验确定了TfR的排除体积，并发现在其理论值的误差范围内。本文的结果表明，SSN在单体和二聚体分子共存于异质混合物中的情况下，成功地应用于准确分类。
A nanopore device is capable of providing single‐molecule level information of an analyte as they translocate through the sensing aperture—a nanometer‐sized through‐hole—under the influence of an applied electric field. In this study, a silicon nitride (SixNy)‐based nanopore was used to characterize the human serum transferrin receptor protein (TfR) under various applied voltages. The presence of dimeric forms of TfR was found to decrease exponentially as the applied electric field increased. Further analysis of monomeric TfR also revealed that its unfolding behaviors were positively dependent on the applied voltage. Furthermore, a comparison between the data of monomeric TfR and its ligand protein, human serum transferrin (hSTf), showed that these two protein populations, despite their nearly identical molecular weights, could be distinguished from each other by means of a solid‐state nanopore (SSN). Lastly, the excluded volumes of TfR were experimentally determined at each voltage and were found to be within error of their theoretical values. The results herein demonstrate the successful application of an SSN for accurately classifying monomeric and dimeric molecules while the two populations coexist in a heterogeneous mixture.
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