Protein-mediated transport across hybrid lipid-block copolymer membranes
Professor Lars Jeuken, University of Leeds
“It was fortunate to get this scholarship, taken up by one of our most talented undergraduate students. The project was challenging, but ultimately successful, providing valuable pilot data to guide our future research in biotechnology”
Prof Lars Jeuken, University of Leeds
Membrane proteins have a wide range of potential applications in industry. However, their application can be limited by the short functional lifetime of expressed proteins, particularly when reconstituted into liposomes.
Previous research has shown that reconstituting membrane proteins in hybrid lipid-block copolymer vesicles can increase their functional half-life by an order of magnitude. Building on this, the challenge is to develop new methodologies for the purpose of enhancing the stability and/or longevity of membrane proteins.
Lars Jeuken is a Professor of Molecular Biophysics at the University of Leeds. The research in his laboratory aims to developed novel biophysical and biotechnological tools for membrane proteins.
Professor Jeuken applied for a CBMNet Vacation Scholarship Award to fund an undergraduate student to carry out research in his laboratory. The project aimed to develop new methodologies to create hybrid polymer-lipid-protein vesicles from polymersomes and bacterial inner membrane extracts. The ultimate goal is to enhance the stability and/or longevity of membrane enzymes.
This research project tested various methods to induce the formation of hybrid lipid-polymer vesicles, including tip-sonication, freeze-thaw and destabilisation of polymersomes with Triton X-100. Of these, only detergent-destabilisation with Triton X-100 was successful in forming hybrid vesicles from bacterial inner membrane extracts and polymersomes.
Hybrid vesicles were analysed by density gradient ultracentrifugation, showing that the ratio between polymer and inner membrane was heterogeneous. Importantly, the hybrid vesicles contained an active membrane-bound hydrogenase from the bacterial inner membrane.
In future experiments, it will be important to assess whether hybrid lipid-polymer-inner membrane vesicles are able to enhance the stability of the membrane-bound hydrogenase from the inner membrane extract. It will also be essential to determine whether recombinant proteins reconstituted into hybrid vesicles can orientate and function correctly.
The data generated in this project will be used as pilot data or preliminary data to support a BBSRC response mode proposal. The undergraduate student gained valuable practical laboratory experience in an IB-focused field and, in the future, would like to pursue a PhD in a related area.
“The scholarship I received helped me to develop my skills and gave me insight in a mysterious and wonderful world of science”
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