Single-molecule peptide fingerprinting

Journal article (2018)

Authors

H.G.T.M. van Ginkel BN/Chirlmin Joo Lab - AS , Kavli institute of nanoscience Delft, QN/Kavli Nanolab Delft - AS
M. Filius Kavli institute of nanoscience Delft, BN/Chirlmin Joo Lab - AS , QN/Kavli Nanolab Delft - AS
M. Szczepaniak BN/Chirlmin Joo Lab - AS , QN/Kavli Nanolab Delft - AS
P. Tulinski BN/Chirlmin Joo Lab - AS , Kavli institute of nanoscience Delft
A.S. Meyer Kavli institute of nanoscience Delft, BN/Anne Meyer Lab
C. Joo BN/Chirlmin Joo Lab - AS , Kavli institute of nanoscience Delft
Research Group
BN/Chirlmin Joo Lab (AS) (TU Delft)
ClpXP Single-molecule FRET Peptides Protein analysis Single-molecule protein sequencing
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Published Date

2018

Language

English

Reuse Rights

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Faculty

Applied Sciences

Department

BN/Bionanoscience

Research Group

BN/Chirlmin Joo Lab

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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