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Model-free photon analysis of diffusion-based single-molecule FRET experiments

Author

Listed:
  • Ivan Terterov

    (Weizmann Institute of Science)

  • Daniel Nettels

    (University of Zurich)

  • Tanya Lastiza-Male

    (Weizmann Institute of Science
    University of Illinois, Urbana-Champaign)

  • Kim Bartels

    (Centre for Structural Systems Biology (CSSB) DESY
    European Molecular Biology Laboratory Hamburg
    University Medical Center Hamburg Eppendorf)

  • Christian Löw

    (Centre for Structural Systems Biology (CSSB) DESY
    European Molecular Biology Laboratory Hamburg)

  • Renee Vancraenenbroeck

    (Weizmann Institute of Science
    University College London)

  • Itay Carmel

    (Weizmann Institute of Science)

  • Gabriel Rosenblum

    (Weizmann Institute of Science)

  • Hagen Hofmann

    (Weizmann Institute of Science)

Abstract

Photon-by-photon analysis tools for diffusion-based single-molecule Förster resonance energy transfer (smFRET) experiments often describe protein dynamics with Markov models. However, FRET efficiencies are only projections of the conformational space such that the measured dynamics can appear non-Markovian. Model-free methods to quantify FRET efficiency fluctuations would be desirable in this case. Here, we present such an approach. We determine FRET efficiency correlation functions free of artifacts from the finite length of photon trajectories or the diffusion of molecules through the confocal volume. We show that these functions capture the dynamics of proteins from nano- to milliseconds both in simulation and experiment, which provides a rigorous validation of current model-based analysis approaches.

Suggested Citation

  • Ivan Terterov & Daniel Nettels & Tanya Lastiza-Male & Kim Bartels & Christian Löw & Renee Vancraenenbroeck & Itay Carmel & Gabriel Rosenblum & Hagen Hofmann, 2025. "Model-free photon analysis of diffusion-based single-molecule FRET experiments," Nature Communications, Nature, vol. 16(1), pages 1-16, December.
    • Handle: RePEc:nat:natcom:v:16:y:2025:i:1:d:10.1038_s41467-025-60764-8
    DOI: 10.1038/s41467-025-60764-8
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