Neurodegenerative disorders are characterised by the progressive deterioration and loss of certain neuronal populations. Distinct clinical features are associated with different diseases, but common fundamental processes have been identified. One example of this is injury via oxidative stress. The production of oxidative species is linked to mitochondrial dysfunction and inflammation, which have been demonstrated to occur early in these disorders. Identifying the proteins that are involved in the response to these species is of critical importance to understanding these diseases. This will provide opportunities to identify novel biomarkers and develop treatments that are effective at these early stages.
The use of genetic code expansion to functionalise proteins with clickable amino acids is an emerging protein engineering technology with many potential applications in neurobiology. Recently, it has been demonstrated that any of the 20 natural amino acids can be replaced with a non-canonical amino acid (ncAA) on a proteome-wide scale. However, the application of this technology in complex cells, such as neurons, is still in its infancy. I aim to establish a novel proteome-wide ncAA incorporation methodology to monitor changes in the neuronal proteome in response to oxidative injury. This will be achieved using ncAAs that have been optimised for an inverse-electron-demand Diels-Alder click reaction, which is highly biorthogonal and has extremely rapid kinetics. Adding a clickable biotin handle will facilitate protein enrichment in a highly time-resolved manner. Subsequent proteomic analysis will be applied to identify changes within the whole proteome or subsets of the proteome. Monitoring changes in protein expression in this way will provide valuable information about neurodegenerative disorders and allow for precise investigations of potential new drug targets and biomarkers.
Fields of science
- HORIZON.1.2 - Marie Skłodowska-Curie Actions (MSCA) Main Programme