Project No. 2444

STANDARD PROJECT

Primary Supervisor

Dr Richard Meek – University of Southampton

Co-Supervisor(s)

Prof Andy Pickford- University of Portsmouth

Prof Max Crispin – University of Southampton

Summary

To develop and function brain cells rely on highly complex and intertwined signalling networks.

To develop and function brain cells rely on highly complex and intertwined signalling networks. Regulation of these networks is achieved, in part, through modifying proteins with post-translational modifications e.g. phosphorylation and glycosylation. Glycosylation is the covalent attachment of sugars onto macromolecules by enzymes. In brain, sugar patterns are highly diverse and control unique landscapes of specialised signalling networks. Sugar cycling is controlled by two types of enzymes: glycosyltransferases (GTs) which attach sugars and glycoside hydrolases (GHs) which remove sugars. Mutations in these enzymes severely impact global and tissue-specific developmental processes, and often cause disease. This project will address fundamental biological questions about the role of GTs and GHs in neuronal development and astrocyte physiology. We will:

1) Purify human GTs and GHs from bacterial, mammalian and insect cell expression systems.

2) Use biochemical and biophysical approaches, including isothermal titration calorimetry and mass spectrometry, to understand which macromolecules are targeted by enzymes and the processes by which sugars are installed and removed.

3) Provide atomic-level insights into the mechanism of enzymes through structural biology (X-ray crystallography and cryo-EM).

4) Explore the dynamics of these modifications in cell lines using custom-made chemical and enzymatic tools.

This research will have a broad impact on our understanding of fundamental neurobiology and glycobiology. It will provide intricate details into how GTs and GHs interpret external stimuli and nutrient changes to modify developmental and cell maintenance pathways. Furthermore, the research proposed will enable us, in collaboration with international experts in chemical glycobiology, to design research tools for the scientific community.

The candidate should have an interest in enzymes and cell signalling. This project will provide extensive training in biochemistry, structural biology, and protein purification. The student will also get hands on experience with different cell culturing techniques.