Resveratrol Modulation of Protein Expression in parkin-Mutant Human Skin Fibroblasts: A Proteomic Approach.

Daniele Vergara,Antonio Gaballo,Tiziana Cocco,Michele Maffia,Marco Di Paola,Anna Ferretta,Paola Tanzarella,Consiglia Pacelli,Anna Signorile

doi:10.1155/2017/2198243

Abstract

In this study, we investigated by two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS) analysis the effects of resveratrol treatment on skin primary fibroblasts from a healthy subject and from a parkin-mutant early onset Parkinson's disease patient. Parkin, an E3 ubiquitin ligase, is the most frequently mutated gene in hereditary Parkinson's disease. Functional alteration of parkin leads to impairment of the ubiquitin-proteasome system, resulting in the accumulation of misfolded or aggregated proteins accountable for the neurodegenerative process. The identification of proteins differentially expressed revealed that resveratrol treatment can act on deregulated specific biological process and molecular function such as cellular redox balance and protein homeostasis. In particular, resveratrol was highly effective at restoring the heat-shock protein network and the protein degradation systems. Moreover, resveratrol treatment led to a significant increase in GSH level, reduction of GSSG/GSH ratio, and decrease of reduced free thiol content in patient cells compared to normal fibroblasts. Thus, our findings provide an experimental evidence of the beneficial effects by which resveratrol could contribute to preserve the cellular homeostasis in parkin-mutant fibroblasts.

Highlights

Parkinson’s disease (PD) is a multifactorial neurodegenerative disorder that predominantly affects the population over 65 years of age [1]
We modified some of the experimental parameters used previously to separate proteins from fibroblast cultures, including 2-DE buffer composition and isoelectric focusing conditions, in order to increase the number of proteins separated by 2-DE and the potential number of differentially expressed proteins identified after comparative analysis
We focused on control (CTR) and PD patient (P1) fibroblasts that we recently characterized for a variety of cellular alterations associated with the modulation of metabolic and cytoskeletal proteins [13, 34]

Summary

Introduction

Parkinson’s disease (PD) is a multifactorial neurodegenerative disorder that predominantly affects the population over 65 years of age [1]. Several risk factors were identified including disease-causing mutations in a specific set of genes that mediate the autosomaldominant or autosomal-recessive forms of PD [3], among which mutations in alpha synuclein (SNCA) and in leucinerich repeat kinase 2 (LRRK2) are responsible for autosomaldominant PD forms whereas mutations in parkin, PTENinduced putative kinase 1 (PINK1), DJ-1, and ATP13A2 are accountable for PD that displays an autosomal recessive mode of inheritance [3]. The most common mutant gene implicated in familial PD is parkin, and various loss-of-function mutations occurring in both alleles produce an aggressive, generally early form of PD [4,5,6]. Parkin is a cytosolic protein with E3 ubiquitin ligase activity, for ubiquitin-proteasome-dependent protein turnover, with a central role in mitochondrial maintenance and turnover. In response to mitochondrial damage, PINK1 induces the activation of parkin by phosphorylation

Methods

Results

Conclusion