Primary Supervisor: Professor Roslyn Bill, Life & Health Sciences
Secondary supervisor: Dr Alice Rothnie and Dr John Simms
PhD project title: In-depth structural characterization of the tetraspanin CD81
University of Registration: Aston University
CD81 is a member of the tetraspanin family of membrane proteins that have broad biological and medical importance. CD81 is involved in cell-cell adhesion, cell proliferation, the immune response, fertilization and infection by several important human pathogens including influenza, HIV, the malarial plasmodium parasite and hepatitis C virus (HCV). CD81 forms diverse, incompletely-understood associations with other proteins in cell membranes: the oligomeric status of CD81 within these complexes, the role of its interactions with specific lipids and the mechanistic detail of how CD81 exerts its biological function are not known.
The CD81 crystal structure identifies a cholesterol-binding pocket; cholesterol binding appears to modulate CD81 activity in cells and has been suggested as a potential mechanism for the regulation of tetraspanin function prompting a need to understand the role of the membrane lipid environment in CD81 folding and function.
We have initiated biophysical and molecular modelling studies of recombinant CD81 that has been extracted in either detergent or styrene maleic acid co-polymer. The latter enables us to understand the importance of the lipid context. Our antibody panel will enable us to identify the structural context of native CD81 oligomers, bridging the resolution gap between the structural and cell biology of CD81. Characterizing the structure of CD81 in this depth will establish a framework to address fundamental questions about tetraspanins in health and disease.
- Ayub H, Clare M, Milic I, Chmel NP, Böning H, Devitt A, Krey T, Bill RM, Rothnie AJ: CD81 extracted in SMALP nanodiscs comprises two distinct protein populations within a lipid environment enriched with negatively charged headgroups, Biochim Biophys Acta Biomembr, 2020, 1862(11):183419.
- Grove J, Hu K, Farquhar MF, Goodall M, Jamshad M, Drummer HE, Bill RM, Balfe P and McKeating JA: A new panel of epitope mapped monoclonal antibodies recognising the prototypical tetraspanin CD81, Wellcome Open Res, 2017, 2:82.
BBSRC Strategic Research Priority: Integrated Understanding of Health: Ageing & Understanding the Rules of Life: Structural Biology
Techniques that will be undertaken during the project:
- Cell culture
- Expression and purification of membrane proteins
- Functional assays
- Molecular modelling and simulation
Contact: Professor Roslyn Bill, AstonUniversity