BiLamVesicles · Novel bi-lamellar lipid vesicles for studying double-membrane transenvelope proteins
Horizon 2020 — Marie Skłodowska-Curie Actions
- Duration
- 2020-08-01 → 2022-07-31
- EU contribution
- €224,934
- Participants
- 1
- Scheme
- MSCA-IF
Lines connect the coordinator with its partners.
Results in brief
Novel bi-lamellar lipid vesicles for studying double-membrane transenvelope proteins
Bi-lamellar membranes, composed of two lipid bi-layers, are ubiquitous in cellular organisms with examples ranging from the double membrane envelope of Gram-negative bacteria to the two lipid-bilayers of eukaryotic subcellular organelles like the cell nucleus and mitochondrion. These double-membranes host sophisticated protein systems that perform many key activities which are crucial for the survival of the cell. For instance, bacterial efflux pumps such as AcrAB-TolC in Escherichia coli are perhaps the most important component in the bacterial line of defense against antibiotic treatment; and while molecularly detailed structures of many types of multidrug transporters are now available, the complex nature of double membranes renders the study of these proteins in live cells highly challenging. In addition, contemporary artificial vesicles like liposomes and other artificial membrane models are inappropriate for hosting and studying this important class of proteins as they either composed of a single or multiple lipid bilayers. My postdoctoral research, as a Marie-Curie fellow, aims to develop a novel artificial double-membrane vesicle for studying the permeation and transport of various molecules through a membranous model that mimics the membrane architecture of various bacteria and organelles.
Data: CORDIS, © European Union
Project 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.
Original text from CORDIS.
Participants
- THE CHANCELLOR MASTERS AND SCHOLARS OF THE UNIVERSITY OF CAMBRIDGE · CAMBRIDGECoordinatorUnited Kingdom
Links
Data: CORDIS, © European Union