Proteomic analysis of changes to mitochondrial oxidative phosphorylation complexes in model systems of Parkinson’s disease

Parkinson’s disease (PD) is a highly debilitating, age-related neurodegenerative disease, which affects all populations, with approximately 120,000 confirmed cases in the UK and 1.5 million in the USA. These numbers are expected to rise as the number of people living over 60 years of age increases. In the general population, there is likely to be a large younger population with asymptomatic disease as significant neurodegeneration occurs before clinical symptoms become apparent as a result of the death of dopaminergic neurons. At present, treatment is purely symptomatic; there is no treatment for the underlying cause of the disease.
The aetiology of the disease is not understood. Although the majority (>90%) of disease is not obviously inherited, studies on proteins linked to inherited forms of the disease point to problems with mitochondrial activity in the more common idiopathic forms of the disease. In this regard, we have evidence that -synuclein (the major protein component of Lewy bodies) and Parkin (a ubiquitin-protein ligase), which are proteins associated with both hereditary and sporadic forms of Parkinson’s disease, associate with and modulate mitochondrial activity (Davison et al (2009) Proteomics 9, 4284-4297; Pennington et al (2010) J. Proteome Res. 9, 2390-2401). We will employ various state-of-the-art proteomic approaches to test the hypothesis that they exert their effects by promoting changes to the composition and / or post-translational modification status of individual components of the mitochondrial oxidative phosphorylation complexes. The initial focus will be to study OXPHOS complex 1, which is composed of 46 component proteins.
The student will gain experience of many basic molecular biology (cDNA cloning and expression), biochemical (mitochondrial isolation and functional assays) and proteomic (one and two dimensional gel electrophoretic techniques coupled with mass spectrometry) techniques, as well as cell culture (generation of permanently transfected cell lines) and immunocytochemistry.