Étude des mécanismes de translocation des peptides pénétrateurs de cellules : approche en microfluidique, électrophysiologie et mesures de forces

Abstract : Cell Penetrating Peptides (CPPs) are known to be able to cross cell membranes with a cargo through two different mechanisms: endocytosis and direct translocation. However, the molecular mechanism of the translocation remains unknown. Our objectives are to find the intermediate structures formed by the CPP and the protein, sugar, and lipid partners, to quantify the strength of the interactions and to measure the kinetics of the steps of the interaction of a CPP with a membrane. This is achieved by using two approaches : (i) coupled BioMembrane Force and electrophysiology, (ii) model membrane in inverse emulsions. (i) Biomembrane Force Probe is a technique for measuring the adhesion force between few molecules grafted on a micrometric bead and a target cell or a liposome. This technique has been used to identify partners of CPP at the cell surface. Electrophysiological measurements on the target cell enable to detect a possible perturbation of a membrane interacting with CPPs. (ii) Inverse emulsions are aqueous droplets into oil covered by lipids. At the interface between two adhering droplets a bilayer is formed. We monitor the translocation of fluorescently labeled CPP through this bilayer. The translocation of several CPPs through negatively charged bilayer within tens of minutes has been detected. The nature of the lipids and the asymmetry of the bilayer seem to impact the translocation. We are currently improving the formation of the pairs of droplets with microfluidic devices for statistical analysis.
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Simon Kulifaj. Étude des mécanismes de translocation des peptides pénétrateurs de cellules : approche en microfluidique, électrophysiologie et mesures de forces. Biophysique. Sorbonne Université, 2018. Français. ⟨NNT : 2018SORUS598⟩. ⟨tel-02477963⟩

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