Dresden 2020 – scientific programme

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BP: Fachverband Biologische Physik

BP 9: Poster II

BP 9.16: Poster

Monday, March 16, 2020, 17:30–19:30, P2/2OG

Isolation of Plasma Membrane Lipids from Immobilized Trypanosomes for Model Membrane Studies — •Nicolas Hagedorn and Susanne Fenz — Department for Cell and Developmental Biology, Biocenter, University of Würzburg, Germany

The unicellular parasite Trypanosoma brucei exhibits a dense, but dynamic, homogeneous surface coat of GPI-anchored variant surface glycoproteins (VSGs). However, its plasma membrane is compartmentalized in three structural and functional distinct domains. In contrast to the homogeneous distribution of VSG, proteins with a GPI anchor have been reported to associate with membrane domains. For trypanosomes, the lipid composition of the plasma membrane is still unknown. We propose plasma membrane vesicles (PMVs) to enrich our understanding of the membrane composition and organization. PMVs were prepared by an approach that combines hypotonic swelling followed by hypertonic cell shrinkage. The effects and mechanisms of vesiculation were studied using light-, and electron microscopy. Microscopic evidence suggests that the main vesiculation site was the flagellum. Moreover, the occurrence of VSGs in PMVs was validated by preparation from trypanosomes exhibiting a fluorescently tagged VSG coat. In the future, formation of solid supported lipid bilayers from PMVs will enable us to address the lipid organization as well as distribution and dynamics of VSGs in a model membrane with natural composition using single-molecule fluorescence microscopy.