Single-molecule peptide fingerprinting

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

doi:10.1073/pnas.1707207115

Single-molecule peptide fingerprinting

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

^*Corresponding author for this work

Research output: Contribution to journal › Article › Scientific › peer-review

54 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 language	English
Pages (from-to)	3338-3343
Number of pages	6
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	115
Issue number	13
DOIs	https://doi.org/10.1073/pnas.1707207115
Publication status	Published - 2018

Keywords

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

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10.1073/pnas.1707207115

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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.",

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T1 - Single-molecule peptide fingerprinting

AU - Van Ginkel, Jetty

AU - Filius, Mike

AU - Szczepaniak, Malwina

AU - Tulinski, Pawel

AU - Meyer, Anne S.

AU - Joo, Chirlmin

PY - 2018

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AB - 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.

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KW - Peptides

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KW - Single-molecule FRET

KW - Single-molecule protein sequencing

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Single-molecule peptide fingerprinting

Abstract

Keywords

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