Berlin 2018 – wissenschaftliches Programm
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BP: Fachverband Biologische Physik
BP 23: Bioimaging and Biopspectroscopy II
BP 23.8: Vortrag
Mittwoch, 14. März 2018, 17:00–17:15, H 2013
Investigating Transient Peptide-Membrane Interactions with TIR-FCS — •Philipp Blumhardt1,3, Jonas Mücksch1,3, Henri G. Franquelim1, Maximilian T. Strauss1,2, Philipp Glock1, Johannes Stein1, Ralf Jungmann1,2, and Petra Schwille1 — 1Max Planck Institute for Biochemistry, Martinsried, Germany — 2Ludwig-Maximilians-Universität, Munich, Germany — 3authors contributed equally
The accurate determination of binding rates to membranes or membrane-bound proteins is of key relevance for quantitative biology. Despite the existence of multiple methods to characterize surface interactions, there are still many experimental challenges regarding simplicity of use and general applicability. We developed a simple and versatile single-molecule fluorescence approach for the accurate determination of binding rates to surfaces or surface-bound receptors. Our approach combines Fluorescence Correlation Spectroscopy (FCS) with Total Internal Reflection (TIR) Fluorescence microscopy and a camera-based detection. This combination not only yields a high surface selectivity, but also resolves association and dissociation rates over a wide time range. Previously, we quantified the transient hybridization of single-stranded DNA to the complementary handles of immobilized DNA origamis. We varied the nucleotide overlap, yielding different transient binding times in the range of milliseconds to tens of seconds. Here, we present our latest results on the transfer of this assay to the otherwise challenging quantification of transient interactions between protein segments and lipid bilayers.