Full-Length Single-Molecule Protein Fingerprinting

Mike Filius, Raman van Wee, Carlos de Lannoy, Ilja Westerlaken, Zeshi Li, Sung Hyun Kim, Cecilia de Agrela Pinto, Yunfei Wu, Geert-Jan Boons, Martin Pabst, Dick de Ridder, Chirlmin Joo*

*Corresponding author for this work

Research output: Working paperPreprintAcademic

Abstract

Proteins are the primary functional actors of the cell. Hence, their identification is pivotal to advance our understanding of cell biology and disease. Current protein analysis methods are of limited use for distinguishing proteoforms. In particular, mass spectrometric methods often provide only ambiguous information on post-translational modification sites, and sequences of co-existing modifications may not be resolved. Here we demonstrate FRET-based single-molecule protein fingerprinting to map the location of individual amino acids and a post-translational modification within single full-length protein molecules. Using an approach that relies on transient binding of fluorescently labeled DNA strands to probe the amino acids on a protein one by one we show that we can fingerprint intrinsically disordered proteins as well as folded globular proteins with sub-nanometer resolution. We anticipate that this technology will be used for proteoform identification in biological and translational research with ultimate sensitivity.
Original languageEnglish
PublisherbioRxiv
Number of pages20
DOIs
Publication statusPublished - 28 Sept 2023

Publication series

NamebioRxiv
PublisherCold Spring Harbor Laboratory
ISSN (Print)2692-8205

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