- Dr Mohanan Maharaj (Research Assistant)
- Mr Carson Stephens (Research Associate)
The Protein Ablation Cancer Therapeutics (PACT) laboratory are interested in understanding how post-translational modifications contribute to the tumorigenic functions of proteins in cancer cells. We hypothesise that particular post-translational modifications are required for the cancer-associated function of a protein and that prevention of these would be a useful approach to treating cancer.
The aim of this project is to select a candidate protein from our database of potential targets, confirm the protein is modified, identify the key modified lysine in the proteins amino acid sequence and finally determine what effect prevention of modification at this lysine has on protein function, stability and localisation within the cancer cell and on cancer cell growth and viability.
We have used Mass Spectrometry approaches to develop a database of interesting proteins. You will choose a protein of interest from this database for your study.
Modification will be confirmed using immunoprecipitation and in-house enzymatic assays with recombinant proteins. The coding sequence will be cloned for lentiviral expression of the protein in cell lines. Online prediction programs will be used to identify lysine residues that may be modified and these will be altered using site-directed mutagenesis.
Functional assays used will depend on the candidate protein chosen but may include biochemical modification of recombinant protein (using in-house assays), Western analysis of downstream signalling proteins, cell fractionation, immunoprecipitation and fluorescence microscopy.
The effect of altering protein modification on cancer cells will be explored using morphology, proliferation, migration and viability assays.
It is expected that novel protein modifications of cancer-associated proteins in cancer cells will be identified and characterised for their contribution to the function, protein-protein interactions, cellular localisation and stability of that protein.
Skills and experience
You should have a demonstrated interest in cancer biology and have completed several units in the cell and molecular biotechnology learning progression.
Contact the supervisor for more information.