Single-molecule peptide fingerprinting

Jetty Van Ginkel, Mike Filius, Malwina Szczepaniak, Pawel Tulinski, Anne S. Meyer, Chirlmin Joo

Research output: Contribution to journalArticleScientificpeer-review

22 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)3338-3343
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume115
Issue number13
DOIs
Publication statusPublished - 2018

Keywords

  • ClpXP
  • Peptides
  • Protein analysis
  • Single-molecule FRET
  • Single-molecule protein sequencing

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