Biophysical Society Thematic Meeting | Tutzing 2026

Single-Molecule FRET: The Next 30 Years

Friday Speaker Abstracts

DYNAMIC STRUCTURAL BIOLOGY WITH SMFRET AND FRET NANOSCOPY Claus A.M. Seidel ; 1 Heinrich-Heine-University, Physical Chemistry, Duesseldorf, Germany For the last 30 years, multimodal fluorescence spectroscopy and microscopy with multiparameter detection is providing rich insights with respect to the structure, kinetics and spatial localization of biomolecular systems. smFRET offers nanosecond time resolution to study motions biomolecules from nano- to milliseconds [1,2] with a high distance precision of ≤ 2 Å and accuracy of ≤ 5 Å [3]. If more than two FRET states are present, we introduced parametric relations (FRET-lines) [4,5] between two FRET indicators, the donor fluorescence lifetime and the intensity-based FRET efficiency, facilitating the interpretation of complex kinetic networks. Finally, a novel framework for FRET nanoscopy and Optical Pythagoras was introduced to analyze single layer DNA origamis [6]. We imaged single objects within a few seconds accessing distances from µm down to 5 nm (dye localization precision ≈ 1 nm, distance precision between 2-3 nm between a donor and acceptor dye). To enable dissemination of all results according to the FAIR principle, we introduce the flrCIF data representation, which extends established data standards from the Protein Data Bank and allows for archiving fluorescence aided integrative structures for multiple states together with associated kinetic data on exchange in the PDB-Dev repository [7]. [1] Sisamakis, E. et al.; Meth. Enzymol. 475, 455-514 (2010). doi.org/10.1016/S0076-6879(10)75018-7. [2] Lerner, E. et al; eLife 10, e60416 (2021). doi.org/10.7554/eLife.60416 [3] Agam, G. et al.; Nat. Methods 20, 523ff (2023). doi.org/10.1038/s41592-023-01807-0 [4] Barth, A. et al.; J. Chem. Phys. 156, e141501 (2022). doi.org/10.1063/5.0089134 [5] Opanasyuk, O. et al.; J. Chem. Phys. 156, e031501 (2022). doi.org/10.1063/5.0095754 [6] van der Voort, N.T.M. et al. arXiv:2108.00024v2. doi.org/10.48550/arXiv.2108.00024.[7] Hanke, C. et al.; Nat. Methods 21, 1970ff (2024). doi.org/10.1038/s41592-024-02428-x

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