Berlin 2018 – wissenschaftliches Programm
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BP: Fachverband Biologische Physik
BP 38: Membranes and Vesicles II (joint session BP/CPP)
BP 38.6: Vortrag
Freitag, 16. März 2018, 11:00–11:15, H 2013
FCS analysis of protein mobility on lipid monolayers — •Jonas Mücksch1,2, Alena Khmelinskaia1,2, Franco Conci1, Grzegorz Chwastek1, and Petra Schwille1 — 1Max Planck Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried — 2authors contributed equally
In vitro membrane model systems are used to study complex biological phenomena under controlled unadulterated conditions. Lipid monolayers are particularly suited to study lipid packing in an unbiased manner. To explore the effect of lipid packing on protein mobility, we used miniaturized chambers instead of conventional Langmuir-Blodgett troughs. This assay requires only minute amounts of protein and is ideally suited to be combined with single molecule sensitive fluorescence correlation spectroscopy (FCS) to characterize diffusion dynamics. Our results confirm the influence of lipid packing on lipid mobility and validate the use of FCS as an alternative to conventional surface pressure measurements. Furthermore, we study the effect of lipid density on the diffusion of membrane binding biomolecules, ranging from small peptides to large DNA-based nanostructures. We exploit the sensitivity of FCS to characterize protein interactions with the lipid monolayer in a low concentration regime, which is inaccessible to conventional surface pressure measurements. Finally, we relate our measurements to the characteristic hydrodynamic length of the lipid monolayer. Our work provides a detailed strategy for the conduction of point FCS experiments on lipid monolayers, which is the first step towards extensive studies of protein-monolayer interactions.