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Post-Translational Modification Analysis of VDAC1 in ALS-SOD1 Model Cells Reveals Specific Asparagine and Glutamine Deamidation

Mitochondria from affected tissues of amyotrophic lateral sclerosis (ALS) patients show morphological and biochemical abnormalities. Mitochondrial dysfunction causes oxidative damage and the accumulation of ROS, and represents one of the major triggers of selective death of motor neurons in ALS. We...

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Autores principales: Pittalà, Maria Gaetana Giovanna, Reina, Simona, Cubisino, Salvatore Antonio Maria, Cucina, Annamaria, Formicola, Beatrice, Cunsolo, Vincenzo, Foti, Salvatore, Saletti, Rosaria, Messina, Angela
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7761621/
https://www.ncbi.nlm.nih.gov/pubmed/33276691
http://dx.doi.org/10.3390/antiox9121218
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author Pittalà, Maria Gaetana Giovanna
Reina, Simona
Cubisino, Salvatore Antonio Maria
Cucina, Annamaria
Formicola, Beatrice
Cunsolo, Vincenzo
Foti, Salvatore
Saletti, Rosaria
Messina, Angela
author_facet Pittalà, Maria Gaetana Giovanna
Reina, Simona
Cubisino, Salvatore Antonio Maria
Cucina, Annamaria
Formicola, Beatrice
Cunsolo, Vincenzo
Foti, Salvatore
Saletti, Rosaria
Messina, Angela
author_sort Pittalà, Maria Gaetana Giovanna
collection PubMed
description Mitochondria from affected tissues of amyotrophic lateral sclerosis (ALS) patients show morphological and biochemical abnormalities. Mitochondrial dysfunction causes oxidative damage and the accumulation of ROS, and represents one of the major triggers of selective death of motor neurons in ALS. We aimed to assess whether oxidative stress in ALS induces post-translational modifications (PTMs) in VDAC1, the main protein of the outer mitochondrial membrane and known to interact with SOD1 mutants related to ALS. In this work, specific PTMs of the VDAC1 protein purified by hydroxyapatite from mitochondria of a NSC34 cell line expressing human SOD1G93A, a suitable ALS motor neuron model, were analyzed by tryptic and chymotryptic proteolysis and UHPLC/High-Resolution ESI-MS/MS. We found selective deamidations of asparagine and glutamine of VDAC1 in ALS-related NSC34-SOD1G93A cells but not in NSC34-SOD1WT or NSC34 cells. In addition, we identified differences in the over-oxidation of methionine and cysteines between VDAC1 purified from ALS model or non-ALS NSC34 cells. The specific range of PTMs identified exclusively in VDAC1 from NSC34-SOD1G93A cells but not from NSC34 control lines, suggests the appearance of important changes to the structure of the VDAC1 channel and therefore to the bioenergetics metabolism of ALS motor neurons. Data are available via ProteomeXchange with identifier <PXD022598>.
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spelling pubmed-77616212020-12-26 Post-Translational Modification Analysis of VDAC1 in ALS-SOD1 Model Cells Reveals Specific Asparagine and Glutamine Deamidation Pittalà, Maria Gaetana Giovanna Reina, Simona Cubisino, Salvatore Antonio Maria Cucina, Annamaria Formicola, Beatrice Cunsolo, Vincenzo Foti, Salvatore Saletti, Rosaria Messina, Angela Antioxidants (Basel) Article Mitochondria from affected tissues of amyotrophic lateral sclerosis (ALS) patients show morphological and biochemical abnormalities. Mitochondrial dysfunction causes oxidative damage and the accumulation of ROS, and represents one of the major triggers of selective death of motor neurons in ALS. We aimed to assess whether oxidative stress in ALS induces post-translational modifications (PTMs) in VDAC1, the main protein of the outer mitochondrial membrane and known to interact with SOD1 mutants related to ALS. In this work, specific PTMs of the VDAC1 protein purified by hydroxyapatite from mitochondria of a NSC34 cell line expressing human SOD1G93A, a suitable ALS motor neuron model, were analyzed by tryptic and chymotryptic proteolysis and UHPLC/High-Resolution ESI-MS/MS. We found selective deamidations of asparagine and glutamine of VDAC1 in ALS-related NSC34-SOD1G93A cells but not in NSC34-SOD1WT or NSC34 cells. In addition, we identified differences in the over-oxidation of methionine and cysteines between VDAC1 purified from ALS model or non-ALS NSC34 cells. The specific range of PTMs identified exclusively in VDAC1 from NSC34-SOD1G93A cells but not from NSC34 control lines, suggests the appearance of important changes to the structure of the VDAC1 channel and therefore to the bioenergetics metabolism of ALS motor neurons. Data are available via ProteomeXchange with identifier <PXD022598>. MDPI 2020-12-02 /pmc/articles/PMC7761621/ /pubmed/33276691 http://dx.doi.org/10.3390/antiox9121218 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Pittalà, Maria Gaetana Giovanna
Reina, Simona
Cubisino, Salvatore Antonio Maria
Cucina, Annamaria
Formicola, Beatrice
Cunsolo, Vincenzo
Foti, Salvatore
Saletti, Rosaria
Messina, Angela
Post-Translational Modification Analysis of VDAC1 in ALS-SOD1 Model Cells Reveals Specific Asparagine and Glutamine Deamidation
title Post-Translational Modification Analysis of VDAC1 in ALS-SOD1 Model Cells Reveals Specific Asparagine and Glutamine Deamidation
title_full Post-Translational Modification Analysis of VDAC1 in ALS-SOD1 Model Cells Reveals Specific Asparagine and Glutamine Deamidation
title_fullStr Post-Translational Modification Analysis of VDAC1 in ALS-SOD1 Model Cells Reveals Specific Asparagine and Glutamine Deamidation
title_full_unstemmed Post-Translational Modification Analysis of VDAC1 in ALS-SOD1 Model Cells Reveals Specific Asparagine and Glutamine Deamidation
title_short Post-Translational Modification Analysis of VDAC1 in ALS-SOD1 Model Cells Reveals Specific Asparagine and Glutamine Deamidation
title_sort post-translational modification analysis of vdac1 in als-sod1 model cells reveals specific asparagine and glutamine deamidation
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7761621/
https://www.ncbi.nlm.nih.gov/pubmed/33276691
http://dx.doi.org/10.3390/antiox9121218
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