Abstract
Nanopores are single-molecule sensors used in nucleic acid analysis, whereas their applicability towards full protein identification has yet to be demonstrated. Here, we show that an engineered Fragaceatoxin C nanopore is capable of identifying individual proteins by measuring peptide spectra that are produced from hydrolyzed proteins. Using model proteins, we show that the spectra resulting from nanopore experiments and mass spectrometry share similar profiles, hence allowing protein fingerprinting. The intensity of individual peaks provides information on the concentration of individual peptides, indicating that this approach is quantitative. Our work shows the potential of a low-cost, portable nanopore-based analyzer for protein identification.
Highlights
Nanopores are single-molecule sensors used in nucleic acid analysis, whereas their applicability towards full protein identification has yet to be demonstrated
We showed that the introduction of an aromatic residue in the sensing region of Fragaceatoxin C (FraC)
We show that G13F-FraC-T1 can be used to directly sample proteins that are digested by a protease
Summary
Nanopores are single-molecule sensors used in nucleic acid analysis, whereas their applicability towards full protein identification has yet to be demonstrated. We show that an engineered Fragaceatoxin C nanopore is capable of identifying individual proteins by measuring peptide spectra that are produced from hydrolyzed proteins. We show that the spectra resulting from nanopore experiments and mass spectrometry share similar profiles, allowing protein fingerprinting. Our work shows the potential of a low-cost, portable nanopore-based analyzer for protein identification. Most mass analyzers are large, have a high cost of investment, are expensive to maintain, and require specialized operators to function[2,3,6]. In contrast to mass spectrometry devices, nanopore-based analyzers provide a low-cost and high-throughput platform with the adaptability towards native environments[7,8,9,10]. Peptides are of special interest for protein characterization, as they allow identification analogous to bottom-up MS-based proteomics
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
