Dundee Research Interest Group

In my last blog I was just about to begin my first project and I am very happy to say that the first experiment was a success. Our lab investigates two principle proteins: PINK1 and Parkin, both of which can have mutations that are associated with Parkinson’s Disease. These proteins work together to add small tags (known as ubiquitin) to the mitochondria, the engine of the cell that produces energy. These tags flag the mitochondria for destruction much like putting a demolition notice on a power plant. This process is ongoing in all healthy cells as a mechanism to remove any damaged mitochondria but in Parkinson’s disease the PINK1/Parkin pathway becomes defective, leaving damaged mitochondria unchecked. As a result, patients get an accumulation of damaged mitochondria over time which is toxic to the cell and is one of the contributory factors to neuronal cell death. Think of it as all the power stations across the country becoming damaged and short-circuiting the national grid – we’d have a huge power cut nationwide.

Whilst the removal of damaged mitochondria seems to be the most robustly shown function of the PINK1/Parkin pathway, the lab has found several alternative roles of these two proteins in the cell. One such role is to be able to act on a group of proteins known as Rab proteins. After mitochondrial damage, phosphorylation (the addition of a small chemical group PO3-) of these Rab proteins has been observed. This process is hypothesized to involve the action of PINK1/Parkin. The cellular consequences of the phosphorylation of Rab proteins are not yet fully understood and how changes to these phosphorylation events in Parkinson’s Disease contribute to its pathology has yet to be fully elucidated. My task is to investigate whether phosphorylation of Rabs is independent or dependent on Parkin. This means if I take Parkin away from cells, can the Rab still be phosphorylayed or will this phosphorylation disappear.

I have now gained some preliminary results and am conducting further experiments to verify my findings. This will require learning two more experimental techniques: a PhosTag and Immunoprecipitation. Like western blots, as I mentioned in my last blog, these techniques are fundamental to research and have a wide variety of applications, so gaining the practice with these techniques is going to be invaluable for me in the future. 

This week I have been learning how to run a PhosTag gel with samples that I have created from my own tissue culture. This is the first time I have undertaken an entire workflow from growing the cells, treating them with specific conditions to elicit a certain response, generating samples from these cells that can be used in downstream experiments and then finally carrying out the experiment. Up to this point I have been using samples other people had previously generated, so this has been a hugely satisfying process.

With 6 weeks of lab experience under my belt I am feeling very settled in the lab and am enjoying the work immensely. The days are flying by and it feels great to be planning my own experiments and shaping my own work. I am also very grateful to be continuing to work in this turbulent time and I have all my fingers and toes crossed that the lab can remain open over the coming winter to continue their lifechanging research.