Dopaminergic human neuroblastoma SH-SY5Y cells were stably transformed to increase expression of alpha-synuclein, a Parkinson's disease-related protein. Transformed cells were more resistant to oxidative insults, showing a cytoprotective role of alpha-synuclein. The expression of redox chaperonins (DJ-1, HSP70, and 14-3-3) was evaluated by Western blotting. Expression of alpha-synuclein reduced HSP70 levels even in the presence of dopamine, with a twofold increase of DJ-1 in the absence of oxidants. DJ-1 is significantly reduced by dopamine, and even more by dopamine and Cu(II). Increased alpha-synuclein expression did not affect 14-3-3, although dopamine increased its level by 60% in wild-type cells. alpha-Synuclein not only upregulated DJ-1, but also shifted all DJ-1 forms to a single spot at pI = 5.7 not observed in wild-type cells. Dopamine gradually restored the distribution of DJ-I forms to a situation similar to wildtype cells, with the form at pI = 6.1 progressively enriched under oxidative conditions.

alpha-synuclein protects SH-SY5Y cells from dopamine toxicity

COLAPINTO, MONICA;MOLTENI, MONICA;ROSSETTI, CARLO;FASANO, MAURO
2006

Abstract

Dopaminergic human neuroblastoma SH-SY5Y cells were stably transformed to increase expression of alpha-synuclein, a Parkinson's disease-related protein. Transformed cells were more resistant to oxidative insults, showing a cytoprotective role of alpha-synuclein. The expression of redox chaperonins (DJ-1, HSP70, and 14-3-3) was evaluated by Western blotting. Expression of alpha-synuclein reduced HSP70 levels even in the presence of dopamine, with a twofold increase of DJ-1 in the absence of oxidants. DJ-1 is significantly reduced by dopamine, and even more by dopamine and Cu(II). Increased alpha-synuclein expression did not affect 14-3-3, although dopamine increased its level by 60% in wild-type cells. alpha-Synuclein not only upregulated DJ-1, but also shifted all DJ-1 forms to a single spot at pI = 5.7 not observed in wild-type cells. Dopamine gradually restored the distribution of DJ-I forms to a situation similar to wildtype cells, with the form at pI = 6.1 progressively enriched under oxidative conditions.
14-3-3 protein, alpha-synuclein, DJ-1, dopamine toxicity, HSP70, human neuroblastoma, oxidative stress, Parkinson's disease, two-dimensional electrophoresis
Colapinto, Monica; Mila, S; Giraudo, S; Stefanazzi, P; Molteni, Monica; Rossetti, Carlo; Bergamasco, B; Lopiano, L; Fasano, Mauro
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11383/1499743
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