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FraC nanopores with adjustable diameter identify the mass of opposite-charge peptides with 44 dalton resolution

A high throughput single-molecule method for identifying peptides and sequencing proteins based on nanopores could reduce costs and increase speeds of sequencing, allow the fabrication of portable home-diagnostic devices, and permit the characterization of low abundance proteins and heterogeneity in...

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Detalles Bibliográficos
Autores principales: Huang, Gang, Voet, Arnout, Maglia, Giovanni
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6381162/
https://www.ncbi.nlm.nih.gov/pubmed/30783102
http://dx.doi.org/10.1038/s41467-019-08761-6
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author Huang, Gang
Voet, Arnout
Maglia, Giovanni
author_facet Huang, Gang
Voet, Arnout
Maglia, Giovanni
author_sort Huang, Gang
collection PubMed
description A high throughput single-molecule method for identifying peptides and sequencing proteins based on nanopores could reduce costs and increase speeds of sequencing, allow the fabrication of portable home-diagnostic devices, and permit the characterization of low abundance proteins and heterogeneity in post-translational modifications. Here we engineer the size of Fragaceatoxin C (FraC) biological nanopore to allow the analysis of a wide range of peptide lengths. Ionic blockades through engineered nanopores distinguish a variety of peptides, including two peptides differing only by the substitution of alanine with glutamate. We also find that at pH 3.8 the depth of the peptide current blockades scales with the mass of the peptides irrespectively of the chemical composition of the analyte. Hence, this work shows that FraC nanopores allow direct readout of the mass of single peptide in solution, which is a crucial step towards the developing of a real-time and single-molecule protein sequencing device.
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spelling pubmed-63811622019-02-21 FraC nanopores with adjustable diameter identify the mass of opposite-charge peptides with 44 dalton resolution Huang, Gang Voet, Arnout Maglia, Giovanni Nat Commun Article A high throughput single-molecule method for identifying peptides and sequencing proteins based on nanopores could reduce costs and increase speeds of sequencing, allow the fabrication of portable home-diagnostic devices, and permit the characterization of low abundance proteins and heterogeneity in post-translational modifications. Here we engineer the size of Fragaceatoxin C (FraC) biological nanopore to allow the analysis of a wide range of peptide lengths. Ionic blockades through engineered nanopores distinguish a variety of peptides, including two peptides differing only by the substitution of alanine with glutamate. We also find that at pH 3.8 the depth of the peptide current blockades scales with the mass of the peptides irrespectively of the chemical composition of the analyte. Hence, this work shows that FraC nanopores allow direct readout of the mass of single peptide in solution, which is a crucial step towards the developing of a real-time and single-molecule protein sequencing device. Nature Publishing Group UK 2019-02-19 /pmc/articles/PMC6381162/ /pubmed/30783102 http://dx.doi.org/10.1038/s41467-019-08761-6 Text en © The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Huang, Gang
Voet, Arnout
Maglia, Giovanni
FraC nanopores with adjustable diameter identify the mass of opposite-charge peptides with 44 dalton resolution
title FraC nanopores with adjustable diameter identify the mass of opposite-charge peptides with 44 dalton resolution
title_full FraC nanopores with adjustable diameter identify the mass of opposite-charge peptides with 44 dalton resolution
title_fullStr FraC nanopores with adjustable diameter identify the mass of opposite-charge peptides with 44 dalton resolution
title_full_unstemmed FraC nanopores with adjustable diameter identify the mass of opposite-charge peptides with 44 dalton resolution
title_short FraC nanopores with adjustable diameter identify the mass of opposite-charge peptides with 44 dalton resolution
title_sort frac nanopores with adjustable diameter identify the mass of opposite-charge peptides with 44 dalton resolution
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6381162/
https://www.ncbi.nlm.nih.gov/pubmed/30783102
http://dx.doi.org/10.1038/s41467-019-08761-6
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