주철민 (Delft University of Technology)

한빛사논문

조회409

Delft University of Technology

CV File 170 kb

Nat. Nanotechnol., Feb 13 2024, | https://doi.org/10.1038/s41565-023-01598-7 Full-length single-molecule protein fingerprinting

Mike Filius ¹, Raman van Wee ^1,7, Carlos de Lannoy^1,2,7, Ilja Westerlaken¹, Zeshi Li ¹, Sung Hyun Kim ^1,3, Cecilia de Agrela Pinto¹, Yunfei Wu⁴, Geert-Jan Boons^4,5, Martin Pabst ⁶, Dick de Ridder² & Chirlmin Joo ^1,3

¹Department of Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, Delft, The Netherlands.

²Bioinformatics Group, Wageningen University, Wageningen, The Netherlands.

³Department of Physics, Ewha Womans University, Seoul, Republic of Korea.

⁴Department of Chemical Biology and Drug Discovery, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands.

⁵Bijvoet Center for Biomolecular Research, Utrecht University, Utrecht, The Netherlands.

⁶Department of Biotechnology, Delft University of Technology, Delft, The Netherlands.

⁷These authors contributed equally: Raman van Wee, Carlos de Lannoy.

Corresponding author : Correspondence to Chirlmin Joo.

Abstract

Proteins are the primary functional actors of the cell. While proteoform diversity is known to be highly biologically relevant, current protein analysis methods are of limited use for distinguishing proteoforms. Mass spectrometric methods, in particular, often provide only ambiguous information on post-translational modification sites, and sequences of co-existing modifications may not be resolved. Here we demonstrate fluorescence resonance energy transfer (FRET)-based single-molecule protein fingerprinting to map the location of individual amino acids and post-translational modifications within single full-length protein molecules. Our data show that both intrinsically disordered proteins and folded globular proteins can be fingerprinted with a subnanometer resolution, achieved by probing the amino acids one by one using single-molecule FRET via DNA exchange. This capability was demonstrated through the analysis of alpha-synuclein, an intrinsically disordered protein, by accurately quantifying isoforms in mixtures using a machine learning classifier, and by determining the locations of two O-GlcNAc moieties. Furthermore, we demonstrate fingerprinting of the globular proteins Bcl-2-like protein 1, procalcitonin and S100A9. We anticipate that our ability to perform proteoform identification with the ultimate sensitivity may unlock exciting new venues in proteomics research and biomarker-based diagnosis.

논문정보

형식Research article
게재일2024년 02월 (BRIC 등록일 2024-02-15)
연구진국내(교신)+국외 연구진
분야 생명과학 > 생물공학

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