Flippable ELP-decorated proteoliposomes to prepare nano-bioreactors

PI:  Christophe Tribet

Pole of Physical and Biological Chemistry of Living Matter at the Ecole Normale Supérieur

1-year contract

The project will explore a generic route to prepare biocatalytic proteoliposomes nanoreactors via the proliferation of bacterial membranes and temperature-triggered loading of chimeric enzymes. The objective is to design biomimetic active compartments with markedly higher loading capacities as compare to presently “artificial” reconstitution methods.[1,2]  Our approach is based on the development of chimeric membrane-proliferating protein scaffolds displaying Elastic-Like Protein sequences (ELPs) that are temperature-responsive.[3] We propose to combine T-switchable capture of enzymes of interest with the flip of membrane curvature enabling to encapsulate enzymes into functional proteoliposomes. In particular, optimized sequences will enable to explore 1) the effect on curvature and tropism for curved areas of the ELP-fused membrane proteins, i.e. their impact on inter-leaflet excess area, or their partition between flat areas and nanotubes, 2) T-switchable capture of cognate ELP-fused enzymes, 3) and finally inside/out flipping of the liposomes upon application of a stress to achieve encapsulation.

[1, 2] A. Etemadi, et al., Artif. Cells, Nanomedicine, Biotechnol. 2016, 44, 381–391 ; T. Trantidou, et al., Engineering Compartmentalized Biomimetic Micro- and Nanocontainers, Acs Nano 11(7) (2017) 6549-6565.

[3] Royes J., Ilioaia, O.; Lubart, Q.; Angius, F.; Dubacheva, G. V.; Bally, M.; Miroux, B.; Tribet, C., Bacteria-Based Production of Thiol-Clickable, Genetically Encoded Lipid Nanovesicles. Angewandte Chemie International Edition, 2019. 58(22): p. 7395-7399.