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Novel bi-lamellar lipid vesicles for studying double-membrane transenvelope proteins

Project description

Artificial lipid vesicles for studying membrane protein activity

Cell membranes are paramount to cell integrity and support essential cellular activities through their proteins. However, studying the activity of transmembrane proteins has been hampered by the lack of suitable models that can host them. The EU-funded BiLamVesicles project proposes to employ microfluidics to develop highly regulated lipid vesicles that will accommodate transmembrane proteins. The key objective is to study the activity of drug efflux pumps in Gram-negative bacteria. Apart from offering fundamental insight into key biological processes, the resultant technique is expected to revolutionise biotechnology applications and pave the way for new drug screening assays.

Objective

Double membranes are ubiquitous throughout the domains of life, accommodating remarkable protein machineries which are fundamental to the cellular activity. However, the study of these proteins is restricted by the lack of a suitable membrane model to accommodate them. Within the framework of BiLamVesicles I will develop a novel bi-lamellar lipid vesicle as a tool for hosting and studying proteins which naturally span across double membranes such as the nucleus and Gram-negative bacteria envelopes. To integrate the protein of choice within the vesicle envelope I will design and employ a highly regulated layer-by-layer assembly in a microfluidic chip. This approach will combine the host’s expertise in microfluidics and biophysics with my expertise in surface interactions and surface chemistry to allow an exquisite control over the membrane composition of bi-lamellar vesicles and the protein insertion process. Once assembled, I will use these vesicles to study the activity of the entire Gram-negative bacterial transporter system AcrAB-TolC, an archetype multidrug efflux pump of Escherichia coli. I will spatially isolate vesicles in a microfluidic chip and directly quantify transport rates through a full efflux pump system at the single-vesicle-level for the first time, using an advanced optofluidic system. The synergy between microfluidics and the proposed double membrane vesicles will produce a ground-breaking biotechnological technique for studying the activity of as yet inaccessible proteins in a biologically-relevant environment. This research will stretch the existing boundaries set by current membrane models and will pave the way for developing advanced techniques for drug screening assays.

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MSCA-IF - Marie Skłodowska-Curie Individual Fellowships (IF)

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Call for proposal

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(opens in new window) H2020-MSCA-IF-2019

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Coordinator

THE CHANCELLOR MASTERS AND SCHOLARS OF THE UNIVERSITY OF CAMBRIDGE
Net EU contribution

Net EU financial contribution. The sum of money that the participant receives, deducted by the EU contribution to its linked third party. It considers the distribution of the EU financial contribution between direct beneficiaries of the project and other types of participants, like third-party participants.

€ 224 933,76
Address
TRINITY LANE THE OLD SCHOOLS
CB2 1TN CAMBRIDGE
United Kingdom

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Region
East of England East Anglia Cambridgeshire CC
Activity type
Higher or Secondary Education Establishments
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Total cost

The total costs incurred by this organisation to participate in the project, including direct and indirect costs. This amount is a subset of the overall project budget.

€ 224 933,76
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