Understanding membrane fusion at the molecular level using a biomimetic model system

Samenvatting

I propose to design a biomimetic model system able to induce for the first time in-vivo membrane fusion and study its mechanism.

Fusion of lipid bilayers in cells facilitates the active transport of chemicals. Non-viral membrane fusion is regulated by a cascade of proteins as the process is highly regulated both in space and time. In eukaryotic cells, the so-called SNARE protein complex is at the heart of fusion. According to 2013 Nobel Prize laureate Sűdhof, how SNARE proteins promote fusion remains a major question in cell biology. This is because SNAREs are large, complex, membrane bound and therefore hard to handle, to modify and to manipulate. Inspired by SNARE-mediated fusion, I will develop a model system able to in-vivo induce fusion.

The model system consists of a complementary pair of lipidated peptides able to form a heterodimeric coiled coil motif at the membrane interface similar to natural SNAREs. The mechanism of membrane fusion will be studied using biophysical techniques, with an emphasis on peptide-peptide and peptide-lipid interactions. Furthermore, membrane fusion between liposomes and live cells will be studied using optimized model systems. Here, it will be a major challenge to achieve efficient and targeted liposome transport directly into the cytoplasm of cells, circumventing endocytotic pathways. The ultimate goal of this project is to achieve targeted in-vivo fusion of liposomes in zebrafish controlled by the biomimetic model system. Future applications are foreseen in drug delivery and membrane engineering.

The original aspect are:
-Design of fusogenic coiled coil peptides
-Understand coiled coil mediated fusion at the molecular level
-Achieve in-vivo fusion using a biomimetic model system for the first time
-Direct delivery in cytoplasm of cells, circumventing endocytosis pathways
-Coiled coil mediated cell-cell fusion to produce hybridoma cells
-Coiled coils targeting cancer cells in order to achieve membrane fusion selectivity

Kenmerken

Projectnummer

724.014.001

Hoofdaanvrager

Prof. dr. A. Kros

Verbonden aan

Universiteit Leiden, Faculteit der Wiskunde en Natuurwetenschappen, Leiden Institute of Chemistry

Uitvoerders

MG Arias Alpizar MSc, Dr. ir. J. Bussmann, Dr. F. Campbell, A. Ciaffoni MSc, N.S.A. Crone MSc, T.M. van Leeuwe MSc

Looptijd

16/11/2015 tot 31/08/2020