Use of a solid‐state nanopore for profiling the transferrin receptor protein and distinguishing between transferrin receptor and its ligand protein. Issue 1 (1st December 2022)

• Record Type: Journal Article
• Authors: O'Donohue, Matthew; Saharia, Jugal; Bandara, Nuwan; Alexandrakis, Georgios; Kim, Min Jun
• Other Names: Lapizco‐Encinas Blanca guestEditor.; Wätzig Hermann guestEditor.
• Is Part Of: Electrophoresis. Volume 44:Issue 1/2(2023)
• Page Start: 349
• Page End: 359
• Publication Date: 2022-12-01
• Subjects: dielectric breakdown -- human serum transferrin -- nanopore -- protein unfolding -- transferrin receptor protein; Electrophoresis -- Periodicals; Electrophoresis -- Periodicals; 541.372
• DOI: 10.1002/elps.202200147
• Languages: English
• ISSNs: 0173-0835

Abstract

Abstract: 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 (Si x N y )‐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.