Regensburg 2022 – wissenschaftliches Programm

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BP: Fachverband Biologische Physik

BP 7: Poster 1

BP 7.4: Poster

Montag, 5. September 2022, 18:00–20:00, P1

Towards Advanced Single Particle Tracking of Molecular Motors by Quantum Dot Labeling and Monitoring of the Cytoskeletal Environment — •Paulina Blair, Adrian Lentz, Xiaoyue Shang, Daniel Kuckla, Philipp Hagemann, and Cornelia Monzel — Experimental Medical Physics, Heinrich-Heine Universität, Düsseldorf, Germany

The molecular motor Kif5C plays a central role in the intracellular transport and synaptic transmission of neuronal cells. To understand how Kif5C mediated transport depends on its environment, it is essential to track the motor in the cytoplasmic context. We aim to advance the single molecule tracking of Kif5C by Quantum Dot labeling as well as by characterising the cytoskeletal environment. Quantum Dots are nanoscale semiconductors, which in contrast to genetically encoded fluorophores, offer tracking at enhanced spatio-temporal resolution and over long time scales. In this project we express Kif5C genetically fused to a streptavidin molecule in live cells. Quantum Dots functionalised with biotin are added to the cell sample to bind via streptavidin-biotin interaction to Kif5C. Molecular motors are recorded with millisecond resolution and are analysed using a global Linear Assignment Approach. The microtubule is stained using a silicon rhodamine-based fluorophore. We then derive parameters such as network density or intersections to correlate it with the motor protein dynamics.

Our data will provide insights on the benefit of Quantum Dot labeling, on molecular motor gaits such directed, superdiffusive and subdiffusive motion as well as on effects of the cytoskeletal environment.