Characteristics of DJ-1 protein in
Parkinson’s Disease (PD)
• Neurodegenerative disorder
• Selective loss of dopamine (DA) neurons in the
• Symptoms: resting tremor, rigidity, and slowness of
• Traditional treatment includes substituting for the lack
of DA, eg with L-Dopa. Problematic
• About 1-2% of the population over 65 affected, of these
• 10 different loci have been identified to date whose
mutation appears connected to PD eg alpha-synuclein
• PARK7 locus codes for DJ-1.
• 189 AA protein belonging to the ThiJ/PfpI protease family
• implicated for example in male fertility, cancer etc.
• DJ-1 is widely expressed especially in subcortical regions, in
both neurones and astrocytes.
• Mostly localises to the cytoplasm, but also to mitochondria,
especially in the case of increased oxidative stress.
• In the presence of reactive oxygen species (ROS), the
isoelectric point (pI) of DJ-1 becomes markedly more acidic.
• Sensitive to oxidative stress, mainly through Cysteine 106.
green and purple
Mutations in DJ-1
• Mutations in the DJ-1 gene have been shown to cause
early onset recessive autosomal early onset PD
• L166P point mutation prevents dimerization of DJ-1.
• Other mutations also present, eg early truncation due
to wrong insertion of stop codon.
• One possible cause for PD might be damage of
neurones through inability to deal with the ROS
accumulating during the metabolism of DA.
• Overexpression of DJ-1 has neuroprotective properties
against oxidative stress.
Aims of the project
• Aim is to further characterise DJ-1 and to examine putative
differences between distribution and biochemistry of DJ-1
between PD and healthy brains,
• Investigate DJ-1’s neuroprotective effects in the healthy and
• The project will be based on ongoing studies in the
STUDYING THE BIOCHEMICAL NATURE
OF DJ-1 IN HUMAN BRAIN
- Comparison of insoluble and soluble DJ-1 in different regions of
the brain, eg cerebellum, cortex, striatum, substantia nigra of
healthy and PD brains, as well as sub-cellular localisation, using
2D gel electrophoresis.
- In 2D gel electrophoresis proteins are separated according to
molecular weight (size) and pI.
- DJ-1 also present in glial inclusions in multiple
- But to date it could not be shown that DJ-1
stains Lewy bodies, inclusions abundant in PD.
- Labelling brain slices of various regions,
investigate whether DJ-1 can be localised to
Lewy bodies after all.
- Use antibody against DJ-1, make that antibody
visible with dyes.
- Attempt to co-immunoprecipitate DJ-1 and
CELL CULTURE WORK
• Using various toxins on cell culture in different
concentrations. Eg: rotenone or paraquat
• Investigate how those toxins affect the sub-
cellular localisation of DJ-1.
• Also see how mitochondria are affected by
assaying citrate synthase activity.
• Bandopadhyay et al (2003). The expression of DJ-1 (PARK7) in normal human
CNS and idiopathic Parkinson’s disease. Brain 127: 420-430.
• Bonifati et al (2004). Linking DJ-1 to neurodegeneration offers novel insights for
understanding the pathogenesis of Parkinson’s Disease. J. Mol. Med. 82:
• Canet-Avilés et al (2004). The Parkinson's disease protein DJ-1 is
neuroprotective due to cysteine-sulfinic acid-driven mitochondrial localization.
PNAS 101: 9103-9108.
• Choi et al (2006). Oxidative Damage of DJ-1 is linked to sporadic Parkinson and
Alzheimer Diseases. J. Bio. Chem. 281: 10816-10824.
• Cookson (2005) The Biochemistry of Parkinson’s disease. Annu. Rev. Biochem
• Moore et al (2005). Molecular Pathophysiology of Parkinson’s Disease. Annu.
Rev. Neurosci. 28: 57-87.
• Wilson et al (2003). The 1.1A resolution crystal structure of DJ-1, the protein
mutated in autosomal recessive early onset Parkinson’s disease. PNAS 100:
• Wood-Kaczmar et al (2006). Understanding the molecular causes of Parkinson’s
Disease. Trends in Mol. Med. 388.