Single-molecule peptide fingerprinting

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 115; no. 13; pp. 3338 - 3343
• Main Authors: van Ginkel, Jetty, Filius, Mike, Szczepaniak, Malwina, Tulinski, Pawel, S. Meyer, Anne, Joo, Chirlmin, 주철민
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences 27.03.2018
• Subjects: Amino acids; Biological Sciences; Fingerprinting; Fluorescence resonance energy transfer; Mass spectrometry; Mass spectroscopy; Peptide mapping; Peptides; Proteins; Proteomics; Substrates; single-molecule FRET; peptides; ClpXP; protein analysis; single-molecule protein sequencing
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.1707207115

Proteomic analyses provide essential information on molecular pathways of cellular systems and the state of a living organism. Mass spectrometry is currently the first choice for proteomic analysis. However, the requirement for a large amount of sample renders a small-scale proteomics study challenging. Here, we demonstrate a proof of concept of single-molecule FRET-based protein fingerprinting. We harnessed the AAA+ protease ClpXP to scan peptides. By using donor fluorophore-labeled ClpP, we sequentially read out FRET signals from acceptor-labeled amino acids of peptides. The repurposed ClpXP exhibits unidirectional processing with high processivity and has the potential to detect low-abundance proteins. Our technique is a promising approach for sequencing protein substrates using a small amount of sample.