Section of Ophthalmology and Neuroscience

Philip Robinson

PARKINSON’S DISEASE RESEACH GROUP

Reader in Molecular Biology

p.a.robinson@leeds.ac.uk

Linking abnormal protein processing and mitochondrial function in Parkinson's Disease

Our aim is to determine the molecular basis of Parkinson’s disease. A number of proteins linked to hereditary forms of PD are either linked to problems of handling misfolded and/or post-translationally damaged proteins (e.g. Parkin (a ubiquitin-protein ligase) and alpha-synuclein) or mitochondrial activity (e.g. DJ1 or PINK1). Recent work using animal models suggests that the two processes are linked. We are using proteomic approaches to identify the changes in protein expression patterns caused by these proteins that may affect mitochondrial activity. We have been employing cell based models and a combination of proteomic procedures, which includes Tandem affinity purification (TAP), 2D-PAGE, or 1D-PAGE together with Liquid chromatography-tandem mass spectrometry, to: (i) capture and identify interactants of Parkin and alpha-synuclein; (ii) to identify proteins whose expression levels are modulated in cells by Parkin or alpha-synuclein; and (iii) to determine the effects of alpha-synuclein overexpression on mitochondrial activity. This work is performed in collaboration with the group of Prof. Roz Banks (LIMM). Novel interactions are currently being confirmed and their biological relevance investigated. Additionally, we have initiated a patient-based study to identify biomarkers of Parkinson’s disease in sera or plasma using SELDI based approaches in collaboration with Prof. Roz Banks and Dr. Stuart Jamieson (Consultant Neurologist, Leeds General Infirmary).

Figure 1 2D Proteomic analysis of protein extracts from cells expressing high or low levels of Parkin

Figure 2 Tandem affinity purification of Parkin interactant proteins