- Opens: Friday 11 December 2020
- Number of places: 1
OverviewCardiovascular disease Diabetes Obesity Membrane transport Membrane trafficking
This project is suitable for:
- Fully-Funded Students
- Joint Supervision with International Institutions or places of work
- PhD Plus Programme
Candidates will have a good degree in a biological science-based subject (e.g. biochemistry, biology, cell biolgy)
Insulin stimulates glucose transport into fat and muscle by the regulated trafficking of the facilitative glucose transporter GLUT4 from insulin-sensitive intracellular stores to the plasma membrane (PM).
This insulin-regulated delivery of GLUT4 to the PM is dysfunctional in patients with insulin-resistance and Type-2 diabetes and GLUT4 sorting and/or trafficking is impaired in these patients. Therefore, there is a need to identify the machinery which regulates the sorting of GLUT4.
Scientists have been grappling with the problem of identifying protein interactions for many years. Because interactions are often short-lived and of low affinity, traditional approaches such as yeast two-hybrid screens and co-immunoprecipitations have not always provided a clear picture of complex macromolecular assemblies. GLUT4 is no different: despite many years of effort, proteins that directly bind to GLUT4 have proven elusive. We hypothesise that such proteins must exist as clear domains within the cytosolic portions of GLUT4 are required for correct intracellular sorting, ergo these domains must bind to proteins. We shall use a new approach to identify these proteins.
To study how spatially compartmentalized protein networks assemble into functionally integrated macromolecular complexes, a class of methods termed ‘proximity labeling’ has seen a surge in growth and utility. Proximity ligation uses engineered enzymes (based on biotin ligases) fused to proteins of interest to selectively and covalently tag neighbouring proteins with biotin in living cells. After cell lysis, biotinylated proteins are captured by interaction with streptavidin beads and characterised using Mass Spec approaches. We shall use this approach, coupled to cell and molecular biology to identify and study key proteins that regulate GLUT4 trafficking in fat cells.
S.Morris, N.D.Geoghegan, J.B.A.Sadler, A.M. Koester, H.L.Black, M.Laub, L.Miller, L.F.Heffernan, J.C.Simpson, C.C.Mastick, J.Cooper, N.Gadegaard, N. J.Bryant and G.W.Gould. (2020) PeerJ 8: e8751. https://doi.org/10.7717/peerj.8751 “Characterisation of GLUT4 trafficking in HeLa cells: comparable kinetics and orthologous trafficking mechanisms to 3T3-L1 adipocytes.”
J.B.A.Sadler, C.A.Lamb, C.R.Welburn, D.Kioumourtzoglou, I.S.Adamson, G.W.Gould and N.J.Bryant. Scientific Reports (2019) 9: 4710. doi: 10.1038/s41598-019-40596-5. “The deubiquitinating enzyme USP25 binds tankyrase and regulates insulin-stimulated trafficking of the facilitative glucose transporter GLUT4 in adipocytes.”
Han, S., J. Li, and A. Y. Ting. 2018. 'Proximity labeling: spatially resolved proteomic mapping for neurobiology', Curr Opin Neurobiol, 50: 17-23.
In addition to tuition fees, this project will also require a bench fee of £12,000 per annum
Applicants can apply using the University PEGASUS Application System https://www.strath.ac.uk/science/strathclydeinstituteofpharmacybiomedicalsciences/studywithus-postgraduate/phd/