IBEC seminar: Yunuen Avalos

GUVs as biomimetic systems to study pathogenic-related mechanisms of human parasites

Membrane models such as Giant Unilamellar Vesicles (GUVs) are a powerful tool for studying reactions occurring in living cells. The main advantage of using these biomimetic systems is the vast possibility of controlling the conditions such as membrane composition, surrounding media and temperature; therefore, minimizing the complexity occurring in normal circumstances and allowing us to understand the mechanism of the membrane-related reactions.

In this talk I will focus on the use of GUVs for the reconstitution of the Endosomal Sorting Complex Required for Transport (ESCRT) machinery whose transitory nature on living membranes makes it difficult to study via in vivo systems. The ESCRT machinery is involved in important membrane-remodeling processes in eukaryotes such as cytokinesis, virus budding, plasma membrane repair, neuron pruning, etc. In all of these reactions, the ESCRT machinery orchestrates the fusion of buds that forms away from the cytosol, contrary to the process regulated by clathrin-coated vesicles. In protozoan parasites such as Plasmodium facilparum and Entamoeba histolytica (causative agents of Malaria and Amebiasis, respectively), the ESCRT machinery controls processes involved in the pathogenic mechanisms, such as exosome generation for Plasmodium and endosome maturation in Entamoeba. By using GUVs as a model system, the action of the ESCRT machinery from these and other organisms has been successfully reconstituted. The GUVs system allowed us to unveil the assembly sequence and the function of the proteins at the membrane. Moreover, we have recreated the topology occurring in living cells by a femto-injection approach. The results derived from these experiments can be used to find new therapeutic targets for the eradication of these parasitic diseases.

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