Biophysical Society Thematic Meeting | Tutzing 2026

Single-Molecule FRET: The Next 30 Years

Poster Abstracts

21-POS Board 11 TOOLS AND STANDARDS FOR FRET-ASSISTED INTEGRATIVE STRUCTURAL BIOLOGY Christian A. Hanke 1 ; Mykola Dimura 2 ; Brinda Vallat 3,4 ; Claus A. M. Seidel 2 ; 1 University of Stuttgart, 2nd Institute of Physics, Stuttgart, Germany 2 Heinrich Heine University Düsseldorf, Molecular Physical Chemistry, Düsseldorf, Germany 3 Rutgers, The State University of New Jersey, Research Collaboratory for Structural Bioinformatics Protein Data Bank and the Institute for Quantitative Biomedicine, Piscataway, NJ, USA 4 Rutgers, The State University of New Jersey, Cancer Institute of New Jersey, New Brunswick, NJ, USA Förster Resonance Energy Transfer (FRET) provides detailed insights into biomolecular structure, conformational heterogeneity, and dynamics across a wide range of length and time scales. As a source of long-range distance information, FRET is particularly valuable for studying large, flexible, and dynamic biomolecular systems and provides important restraints in integrative structural biology. We present a workflow supporting different stages of FRET-based structural studies. The methodology assists in the design of FRET experiments by suggesting potential labeling positions and FRET pairs. Experimental FRET data can subsequently be used to screen and score sets of structure models, enabling the identification of conformations that best agree with the measurements and facilitating the generation of FRET-assisted integrative structure models (DOI: 10.1038/s41467-020-19023-1).To promote FAIR data practices, reproducibility, data reuse, and interoperability, we also present approaches for the standardized representation and deposition of FRET experiments and derived structural information. The flrCIF data dictionary (DOI: 10.1038/s41592-024-02428-x) provides a comprehensive description of fluorescence experiments associated with structure models, including samples, fluorescent probes, labeling strategies, experimental conditions, data analysis procedures, and structural modeling workflows. Together with PDB-IHM, flrCIF enables the deposition of FRET-derived restraints, associated metadata, and resulting integrative structure models in the Protein Data Bank (PDB). In addition, the framework supports the representation of complex kinetic schemes, facilitating the archival of dynamic information obtained from fluorescence and complementary biophysical approaches. Furthermore, flrCIF provides a foundation for community-driven standards for the description and deposition of FRET data and may serve as a blueprint for future FRET data repositories. By linking experiment design, structural interpretation, and FAIR data deposition, the presented approaches support both the application of FRET in integrative structural biology and the establishment of community standards for sharing and reusing FRET-derived information.

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