Dresden 2020 – wissenschaftliches Programm

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BP: Fachverband Biologische Physik

BP 20: Poster VIII

BP 20.2: Poster

Dienstag, 17. März 2020, 14:00–16:00, P2/4OG

Studying molecular interactions with a combination of microfluidics and FFS — •Eleonora Perego and Sarah Köster — IRP, Georg-August-Universität Göttingen, Germany

Assembly and aggregation of biomolecules into larger complexes are fundamental processes in living organisms. Ordered protein assembly is vital for the organism, as for example the assembly of the cytoskeletal filaments, however sometimes disordered aggregates, which can be toxic as fro example alpha-synuclein fibrils, are also produced. It is fundamental to study both the ordered assembly and the disordered aggregation with high spatial resolution (on a single molecule level) and good temporal precision (in the order of ms) to gain a complete knowledge of these reactions. Here, we combine fluorescence fluctuation spectroscopy (FFS), employed to measure the interactions, with microfluidics to access the temporal information. We focus on studying the ordered assembly of vimentin, an intermediate filament (IF) protein which is part of the cytoskeleton, using a multi-layer microfluidic device that prevents the protein from coming in contact with the channel walls. This type of device also provides a controlled diffusive mixing of assembly buffer and protein solution. Employing FFS in these devices enables us to precisely measure the labelling stoichiometry of the assembling protein, which allow us to follow the very first time steps of vimentin assembly. Our results show that the combination of microfluidics and FFS provides a suitable approach for studying the aggregation of biomolecules in real time, which is important for understanding cellular behavior.