Single-Molecule Peptide Identification Using Fluorescence Blinking Fingerprints.

Autor: Püntener S; Institute of Chemical Sciences and Engineering, Ecole Polytechnique Fédéral de Lausanne, CH-1015 Lausanne, Switzerland.; Department of Chemistry, University of Zurich, CH-8057 Zurich, Switzerland., Rivera-Fuentes P; Institute of Chemical Sciences and Engineering, Ecole Polytechnique Fédéral de Lausanne, CH-1015 Lausanne, Switzerland.; Department of Chemistry, University of Zurich, CH-8057 Zurich, Switzerland.
Jazyk: angličtina
Zdroj: Journal of the American Chemical Society [J Am Chem Soc] 2023 Jan 18; Vol. 145 (2), pp. 1441-1447. Date of Electronic Publication: 2023 Jan 05.
DOI: 10.1021/jacs.2c12561
Abstrakt: The ability to identify peptides with single-molecule sensitivity would lead to next-generation proteomics methods for basic research and clinical applications. Existing single-molecule peptide sequencing methods can read some amino acid sequences, but they are limited in their ability to distinguish between similar amino acids or post-translational modifications. Here, we demonstrate that the fluorescence intermittency of a peptide labeled with a spontaneously blinking fluorophore contains information about the structure of the peptide. Using a deep learning algorithm, this single-molecule blinking pattern can be used to identify the peptide. This method can distinguish between peptides with different sequences, peptides with the same sequence but different phosphorylation patterns, and even peptides that differ only by the presence of epimerized residues. This study builds the foundation for a targeted proteomics method with single-molecule sensitivity.
Databáze: MEDLINE