Berlin 2024 – wissenschaftliches Programm

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BP: Fachverband Biologische Physik

BP 27: Single Molecule Biophysics

BP 27.7: Hauptvortrag

Donnerstag, 21. März 2024, 11:15–11:45, H 0112

Integrative dynamic structural biology with multi-modal fluorescence spectroscopy and nanoscopy: From single molecules to live cells — •Claus Seidel — Heinrich-Heine-University Düsseldorf, Germany

Multimodal fluorescence spectroscopy and microscopy with multiparameter detection provide rich insights on biomolecular systems under ambient / live cell conditions, including spatial, structural and kinetic information. In a comparative single-molecule study, we assessed the accuracy of Förster Resonance Energy Transfer (FRET) measurements. We studied two protein systems with distinct conformational changes and dynamics and obtained an interdye distance precision of smaller than 2 Å and accuracy of smaller than 5 Å. Considering cellular studies, we introduced a framework for quantitative high throughput FRET image spectroscopy. We measured the time-evolution of pairwise homo- or hetero-interactions of the Guanylate binding proteins and the membrane receptor CD95 in live cells with 0.8% fraction precision. In this way, the next level of complexity is achieved by linking structural dynamics of biomolecules with their cellular function and localization.

Keywords: Single molecules; Fluorescence Spectroscopy; FRET; Dynamics; Superresolution microscopy