Serum from Parkinson’s disease patients presents excess of protein halogenation and nitrosylation,...
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POSTER PRESENTATION Open Access Serum from Parkinson’s disease patients presents excess of protein halogenation and nitrosylation, with anomalous nitrosylation of serum a- synuclein as potentially etiological factor Emilio Fernandez 1* , Jose-Manuel Garcia-Moreno 2 , Angel Martin de Pablos 3 , Jose Chacon 1 From Molecular Neurodegeneration: Basic biology and disease pathways Cannes, France. 10-12 September 2013 Background Halogenative and nitrosative stress are two types of oxida- tive stress that have been proposed as pathogenic mechan- isms in Parkinson’s disease (PD). They can be caused by over-stimulation of phagocytes. We hypothesize that maintained phagocyte overstimulation leads to both halo- genative and nitrosative stress in Parkinson’ s disease, which are present in serum and cerebrospinal fluid of patients. These types of oxidative stress could modify pro- teins related to the pathogenesis of Parkinson’s disease. Materials and methods Halogenative and nitrosative stress-induced protein changes in serum and CSF were analyzed in PD patients (n=54) and control subjects without any neurological disor- der (n=40), by using ELISA, western-blotting, mass spectro- metry, culture techniques and immunofluorescence. Results In our lab, it has been detected the presence of halogena- tive stress in serum and, to a lesser extent, cerebrospinal fluid of Parkinsonian patients leading to excess of advanced oxidized protein products or AOPP. On the other hand, nitrosative stress is also present in serum and cerebrospinal fluid of patients with early PD, character- ized by the selective increase of 3-nitrotyrosine proteins other than nitroalbumin and free 3-nitrotyrosine. Nitro- sylation stress is accompanied by modification of the sites of nitrosylation of a-synuclein in patients with early PD, characterized by dominant nitrosylation of tyrosine 125/136 residues. In vitro studies revealed that 60K- filtrated serum from PD patients is cytotoxic for sub- stanta nigra neurons in culture, and it is able to induce proteinaceous aggregates inside these neurons, which are strongly positive to nitro- a-synuclein. Interestingly, cytotoxicity can be blocked by using antibodies aimed to nitrosylated tyrosine-39 of the molecule. Conclusions Since metabolism of advanced oxidized protein products and 3-nitrotyrosine proteins has been associated to phago- cytic overstimulation, this pathological alteration could play a pathogenic role in sporadic PD. Our observations also lead to the hypothesis that serum level of advanced oxidized protein products is a prognostic marker for Par- kinson’ s disease duration, and these oxidized proteins could participate in neuroinflammation. Besides, the eva- luation of nitrosative stress through enhanced levels of 3-nitrotyrosine proteins in serum and cerebrospinal fluid without changes in nitroalbumin, together with the profile of tyrosine nitrosylation of serum a-synuclein character- ized by dominant nitrosylation of Tyr125/136 could serve for diagnosis of sporadic Parkinson’s disease. Nitro-a-synu- clein is a main component of Lewy bodies, hallmarks of the disease, and serum nitro-a-synuclein could represent a pathogenic factor in PD. Finally, we have in vitro evidence that blockade of tyrosine residue 39 of nitro-a-synuclein could abolish cytotoxicity of Parkinsonian serum. Acknowledgements Supported by grants to EFE by Junta de Andalucia (BIO127), and Spanish Ministerio de Sanidad (RETICS, RD06/001/002; RD06/010/1007; Instituto Carlos III, co-financing with FEDER, European Fund for Regional Development). 1 Department of Medical Physiology, University of Seville, Seville, Spain Full list of author information is available at the end of the article Fernandez et al. Molecular Neurodegeneration 2013, 8(Suppl 1):P18 http://www.molecularneurodegeneration.com/content/8/S1/P18 © 2013 Fernandez et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Transcript of Serum from Parkinson’s disease patients presents excess of protein halogenation and nitrosylation,...
POSTER PRESENTATION Open Access
Serum from Parkinson’s disease patients presentsexcess of protein halogenation and nitrosylation,with anomalous nitrosylation of serum a-synuclein as potentially etiological factorEmilio Fernandez1*, Jose-Manuel Garcia-Moreno2, Angel Martin de Pablos3, Jose Chacon1
From Molecular Neurodegeneration: Basic biology and disease pathwaysCannes, France. 10-12 September 2013
BackgroundHalogenative and nitrosative stress are two types of oxida-tive stress that have been proposed as pathogenic mechan-isms in Parkinson’s disease (PD). They can be caused byover-stimulation of phagocytes. We hypothesize thatmaintained phagocyte overstimulation leads to both halo-genative and nitrosative stress in Parkinson’s disease,which are present in serum and cerebrospinal fluid ofpatients. These types of oxidative stress could modify pro-teins related to the pathogenesis of Parkinson’s disease.
Materials and methodsHalogenative and nitrosative stress-induced proteinchanges in serum and CSF were analyzed in PD patients(n=54) and control subjects without any neurological disor-der (n=40), by using ELISA, western-blotting, mass spectro-metry, culture techniques and immunofluorescence.
ResultsIn our lab, it has been detected the presence of halogena-tive stress in serum and, to a lesser extent, cerebrospinalfluid of Parkinsonian patients leading to excess ofadvanced oxidized protein products or AOPP. On theother hand, nitrosative stress is also present in serum andcerebrospinal fluid of patients with early PD, character-ized by the selective increase of 3-nitrotyrosine proteinsother than nitroalbumin and free 3-nitrotyrosine. Nitro-sylation stress is accompanied by modification of thesites of nitrosylation of a-synuclein in patients with earlyPD, characterized by dominant nitrosylation of tyrosine
125/136 residues. In vitro studies revealed that 60K-filtrated serum from PD patients is cytotoxic for sub-stanta nigra neurons in culture, and it is able to induceproteinaceous aggregates inside these neurons, which arestrongly positive to nitro-a-synuclein. Interestingly,cytotoxicity can be blocked by using antibodies aimedto nitrosylated tyrosine-39 of the molecule.
ConclusionsSince metabolism of advanced oxidized protein productsand 3-nitrotyrosine proteins has been associated to phago-cytic overstimulation, this pathological alteration couldplay a pathogenic role in sporadic PD. Our observationsalso lead to the hypothesis that serum level of advancedoxidized protein products is a prognostic marker for Par-kinson’s disease duration, and these oxidized proteinscould participate in neuroinflammation. Besides, the eva-luation of nitrosative stress through enhanced levels of3-nitrotyrosine proteins in serum and cerebrospinal fluidwithout changes in nitroalbumin, together with the profileof tyrosine nitrosylation of serum a-synuclein character-ized by dominant nitrosylation of Tyr125/136 could servefor diagnosis of sporadic Parkinson’s disease. Nitro-a-synu-clein is a main component of Lewy bodies, hallmarks of thedisease, and serum nitro-a-synuclein could represent apathogenic factor in PD. Finally, we have in vitro evidencethat blockade of tyrosine residue 39 of nitro-a-synucleincould abolish cytotoxicity of Parkinsonian serum.
AcknowledgementsSupported by grants to EFE by Junta de Andalucia (BIO127), and SpanishMinisterio de Sanidad (RETICS, RD06/001/002; RD06/010/1007; Instituto CarlosIII, co-financing with FEDER, European Fund for Regional Development).
1Department of Medical Physiology, University of Seville, Seville, SpainFull list of author information is available at the end of the article
Fernandez et al. Molecular Neurodegeneration 2013, 8(Suppl 1):P18http://www.molecularneurodegeneration.com/content/8/S1/P18
© 2013 Fernandez et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the CreativeCommons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, andreproduction in any medium, provided the original work is properly cited.
Authors’ details1Department of Medical Physiology, University of Seville, Seville, Spain.2Hospital Macarena SAS, Seville, Spain. 3Department of Surgery, University ofSeville, Seville, Spain.
Published: 13 September 2013
doi:10.1186/1750-1326-8-S1-P18Cite this article as: Fernandez et al.: Serum from Parkinson’s diseasepatients presents excess of protein halogenation and nitrosylation, withanomalous nitrosylation of serum a-synuclein as potentially etiologicalfactor. Molecular Neurodegeneration 2013 8(Suppl 1):P18.
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Fernandez et al. Molecular Neurodegeneration 2013, 8(Suppl 1):P18http://www.molecularneurodegeneration.com/content/8/S1/P18
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