Wellcome-MRC Institute of Metabolic Science
We are interested in understanding how insulin stimulates glucose transport into fat and muscle cells, and how this process breaks down in disease. Impaired insulin-stimulated glucose transport in muscle and fat is a major contributor to whole body insulin resistance, the key pathophysiological mechanism underlying type 2 diabetes. There is currently no consensus on the molecular basis for impaired insulin responses in these tissues. We aim to address this knowledge gap to identify how the insulin signalling network and GLUT4 trafficking apparatus are altered in insulin resistance.
PhD projects available with our group
Techniques we use
- Cell culture – we use a range of cell lines, including cultured adipocytes and engineered stable cell lines to study protein trafficking, signalling and metabolism.
- Immunofluorescence microscopy – we use high-content, fixed- and live-cell imaging (e.g. confocal, super resolution) to study protein trafficking and cell morphology.
- Flow cytometry – we label cell surface epitopes to study delivery of proteins to the cell surface.
- RUSH – Retention Using Selective Hooks. A cell-based assay that allows us to synchronise protein trafficking through the secretory pathway.
- Metabolomics & metabolic tracing – we use isotopic tracers (e.g. U13C-glucose, deuterium oxide) to study glucose, amino acid, and lipid metabolism.
- Subcellular proteomics – we use techniques like LOPIT-DC (Localisation of Organelle Proteins by Isotope Tagging after Differential Centrifugation) to map the spatial proteome of adipocytes, and identify proteins that move in response to specific stimuli (e.g. insulin).
We also use a range of molecular biology and biochemical techniques including molecular cloning, qPCR, Western blotting, and immunoprecipitation.
Team achievements
Congratulations to Olivia Conway and Joycelyn Tan who presented their work at the IMS-MRL student symposium in July 2022.
Olivia was also selected for an oral presentation and gave an excellent talk at the 2022 Proteomics in Cell Biology and Disease Mechanisms conference (October 2022).
Recent publications
- Trafficking regulator of GLUT4-1 (TRARG1) is a GSK3 substrate. Duan X, Norris DM, Humphrey SJ, Yang P, Cooke KC, Bultitude WP, Parker BL, Conway OJ, Burchfield JG, Krycer JR, Brodsky FM, James DE, Fazakerley DJ. Biochem J. 2022 Jun 17;479(11):1237-1256. doi: 10.1042/BCJ20220153.
- Integrating adipocyte insulin signaling and metabolism in the multi-omics era. Calejman CM, Doxsey WG, Fazakerley DJ, Guertin DA. Trends Biochem Sci. 2022 Jun;47(6):531-546. doi: 10.1016/j.tibs.2022.02.009.
- GLUT4 On the move. Fazakerley DJ, Koumanov F, Holman GD. Biochem J. 2022 Feb 11;479(3):445-462. doi: 10.1042/BCJ20210073.
Our highlight publication
In 2015, we found that Trafficking regulator of GLUT4-1 (TRARG1) regulates insulin-stimulated glucose uptake into fat cells. Here, we report that TRARG1 is a target of the kinase GSK3, providing new insight into how the insulin signalling network controls glucose transport.
Trafficking regulator of GLUT4-1 (TRARG1) is a GSK3 substrate. Duan X, Norris DM, Humphrey SJ, Yang P, Cooke KC, Bultitude WP, Parker BL, Conway OJ, Burchfield JG, Krycer JR, Brodsky FM, James DE, Fazakerley DJ. Biochem J. 2022 Jun 17;479(11):1237-1256. doi: 10.1042/BCJ20220153.
