Project No.2412

PRIORITY PROJECT

Primary Supervisor

Prof Jon Dawson – University of Southampton

Co-Supervisor(s)

Dr Bruce Lichtenstein – University of Portsmouth

Prof Andy Pickford – University of Portsmouth

Dr Seung Lee – University of Southampton

Summary

Glycosylation is the most abundant post translational modification of proteins and is of fundamental importance across biology.

Particularly important are oligosaccharides attached to proteins and cell surfaces. They are involved in cell-cell recognition, signalling, immune response modulation, and host-microbe interactions and are of interest as potential therapeutic agents or as targets for drug development.

Oligosaccharides are, however, hard to synthesise requiring arduous multi-step synthesis procedures due to their structural complexity. In nature, oligosaccharides are synthesised by multiple enzymes arranged in the Golgi apparatus and endoplasmic reticulum in clusters. Substrates diffuse through these clusters to form a target molecule. There have been attempts to mimic such synthetic machinery, typically using flow or microfluidic reactors, but without great success.

In Southampton we have recently applied self-assembling gels formed from nano-sized clay particles to pattern proteins at very high (micron) resolution within a 3D nanoclay-protein gel. Patterning is highly scalable, can be achieved under biocompatible conditions and patterned proteins can remain stable within clay gels for extended periods.

Combined with the well establish ability of clay to catalyse enzymes through immobilisation, this new approach to patterning suggests exciting potential for multienzyme synthesis of oligosaccharides – as well as many other biomolecules – and thus has considerable translational scope for the engineering of scalable bioreactors and sensors. In this project we will seek to apply this technology to pattern enzymes with the goal of mimicking the sequence of enzymatic reactions involved in glycosylation.

The project will involve the application of computational and lab-based assays of enzyme binding, structure and function on clay surfaces, as well as confocal imaging studies of labelled enzymes within nanoclay assemblies. It would be well suited to a student with a background in biochemistry or molecular biophysics.