@article{11eb0d6ef40e4c67a0716d91cb3201a3,
title = "Single-molecule peptide fingerprinting",
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.",
keywords = "ClpXP, Peptides, Protein analysis, Single-molecule FRET, Single-molecule protein sequencing",
author = "{Van Ginkel}, Jetty and Mike Filius and Malwina Szczepaniak and Pawel Tulinski and Meyer, {Anne S.} and Chirlmin Joo",
note = "Funding Information: ACKNOWLEDGMENTS. We thank Marek Noga, Ivo Severins, and Anna Haagsma for technical support; Stanley Chandradoss and Margreet Docter for building the multicolor optical setup; Luuk Loeff and Misha Klein for assisting with data analysis; and Tim Blosser, Laura Restrepo, Margreet Docter, Stephanie Heerema, Noortje de Haan, and Viktorija Globyte for critical reading of the manuscript. ClpX6 expression plasmids were a generous gift from Andreas Martin. ClpP expression plasmids were a generous gift from Tania Baker and Robert Sauer. Titin expression plasmids were a generous gift from Victor Mu{\~n}oz and J{\"o}rg Sch{\"o}nfelder. C.J. and A.S.M. were funded by the Foundation for Fundamental Research on Matter (Grants Projectruimte 12PR3029 and Vrije Programma SMPS). Publisher Copyright: {\textcopyright} 2018 National Academy of Sciences. All Rights Reserved.",
year = "2018",
month = mar,
day = "27",
doi = "10.1073/pnas.1707207115",
language = "English",
volume = "115",
pages = "3338--3343",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
publisher = "National Academy of Sciences",
number = "13",
}