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BP: Fachverband Biologische Physik

BP 12: Poster II

BP 12.2: Poster

Dienstag, 2. April 2019, 14:00–16:00, Poster B2

Studying molecular interactions under flow — •Eleonora Perego and Sarah Köster — Institute for X-Rays physics, University of Göttingen

In recent years the investigation of assembly and aggregation of proteins attracted attention in the scientific community. These processes are the basis of important cellular mechanisms, thus it is important to gain a complete knowledge of the interactions between molecules. Here, we focus on studying the ordered assembly of intermediate filament proteins (IFs), a cytoskeletal component. Although in human more than 70 types of IFs exist, the assembly pathway is common to all of them. The assembly starts with the lateral association of small rod-shaped monomers forming a so called unit length filament (ULF), that is the starting point for the elongation step. We combine photon counting histogram (PCH) to study this process with high spatial resolution, with microfluidics to achieve high temporal precision. We use a multi-layer microfluidics device that prevents the protein from come in contact with the channel walls. This type of device also provides a controlled mixing of assembly buffer and protein solution. Employing PCH on these devices enables us to precisely measure the labelling stoichiometry of the assembling protein, helping us to understand the first steps of vimentin assembly. Our results show that the combination of microfluidics and single molecule fluorescence provides a suitable approach for studying the aggregation of biomolecules in real time, which is important for understanding cellular behaviour.