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SSADH Variants Increase Susceptibility of U87 Cells to Mitochondrial Pro-Oxidant Insult

Succinate semialdehyde dehydrogenase (SSADH) is a mitochondrial enzyme, encoded by ALDH5A1, mainly involved in γ-aminobutyric acid (GABA) catabolism and energy supply of neuronal cells, possibly contributing to antioxidant defense. This study aimed to further investigate the antioxidant role of SSAD...

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Autores principales: Menduti, Giovanna, Vitaliti, Alessandra, Capo, Concetta Rosa, Lettieri-Barbato, Daniele, Aquilano, Katia, Malaspina, Patrizia, Rossi, Luisa
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7352567/
https://www.ncbi.nlm.nih.gov/pubmed/32575506
http://dx.doi.org/10.3390/ijms21124374
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author Menduti, Giovanna
Vitaliti, Alessandra
Capo, Concetta Rosa
Lettieri-Barbato, Daniele
Aquilano, Katia
Malaspina, Patrizia
Rossi, Luisa
author_facet Menduti, Giovanna
Vitaliti, Alessandra
Capo, Concetta Rosa
Lettieri-Barbato, Daniele
Aquilano, Katia
Malaspina, Patrizia
Rossi, Luisa
author_sort Menduti, Giovanna
collection PubMed
description Succinate semialdehyde dehydrogenase (SSADH) is a mitochondrial enzyme, encoded by ALDH5A1, mainly involved in γ-aminobutyric acid (GABA) catabolism and energy supply of neuronal cells, possibly contributing to antioxidant defense. This study aimed to further investigate the antioxidant role of SSADH, and to verify if common SNPs of ALDH5A1 may affect SSADH activity, stability, and mitochondrial function. In this study, we used U87 glioblastoma cells as they represent a glial cell line. These cells were transiently transfected with a cDNA construct simultaneously harboring three SNPs encoding for a triple mutant (TM) SSADH protein (p.G36R/p.H180Y/p.P182L) or with wild type (WT) cDNA. SSADH activity and protein level were measured. Cell viability, lipid peroxidation, mitochondrial morphology, membrane potential (ΔΨ), and protein markers of mitochondrial stress were evaluated upon Paraquat treatment, in TM and WT transfected cells. TM transfected cells show lower SSADH protein content and activity, fragmented mitochondria, higher levels of peroxidized lipids, and altered ΔΨ than WT transfected cells. Upon Paraquat treatment, TM cells show higher cell death, lipid peroxidation, 4-HNE protein adducts, and lower ΔΨ, than WT transfected cells. These results reinforce the hypothesis that SSADH contributes to cellular antioxidant defense; furthermore, common SNPs may produce unstable, less active SSADH, which could per se negatively affect mitochondrial function and, under oxidative stress conditions, fail to protect mitochondria.
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spelling pubmed-73525672020-07-15 SSADH Variants Increase Susceptibility of U87 Cells to Mitochondrial Pro-Oxidant Insult Menduti, Giovanna Vitaliti, Alessandra Capo, Concetta Rosa Lettieri-Barbato, Daniele Aquilano, Katia Malaspina, Patrizia Rossi, Luisa Int J Mol Sci Article Succinate semialdehyde dehydrogenase (SSADH) is a mitochondrial enzyme, encoded by ALDH5A1, mainly involved in γ-aminobutyric acid (GABA) catabolism and energy supply of neuronal cells, possibly contributing to antioxidant defense. This study aimed to further investigate the antioxidant role of SSADH, and to verify if common SNPs of ALDH5A1 may affect SSADH activity, stability, and mitochondrial function. In this study, we used U87 glioblastoma cells as they represent a glial cell line. These cells were transiently transfected with a cDNA construct simultaneously harboring three SNPs encoding for a triple mutant (TM) SSADH protein (p.G36R/p.H180Y/p.P182L) or with wild type (WT) cDNA. SSADH activity and protein level were measured. Cell viability, lipid peroxidation, mitochondrial morphology, membrane potential (ΔΨ), and protein markers of mitochondrial stress were evaluated upon Paraquat treatment, in TM and WT transfected cells. TM transfected cells show lower SSADH protein content and activity, fragmented mitochondria, higher levels of peroxidized lipids, and altered ΔΨ than WT transfected cells. Upon Paraquat treatment, TM cells show higher cell death, lipid peroxidation, 4-HNE protein adducts, and lower ΔΨ, than WT transfected cells. These results reinforce the hypothesis that SSADH contributes to cellular antioxidant defense; furthermore, common SNPs may produce unstable, less active SSADH, which could per se negatively affect mitochondrial function and, under oxidative stress conditions, fail to protect mitochondria. MDPI 2020-06-19 /pmc/articles/PMC7352567/ /pubmed/32575506 http://dx.doi.org/10.3390/ijms21124374 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
Menduti, Giovanna
Vitaliti, Alessandra
Capo, Concetta Rosa
Lettieri-Barbato, Daniele
Aquilano, Katia
Malaspina, Patrizia
Rossi, Luisa
SSADH Variants Increase Susceptibility of U87 Cells to Mitochondrial Pro-Oxidant Insult
title SSADH Variants Increase Susceptibility of U87 Cells to Mitochondrial Pro-Oxidant Insult
title_full SSADH Variants Increase Susceptibility of U87 Cells to Mitochondrial Pro-Oxidant Insult
title_fullStr SSADH Variants Increase Susceptibility of U87 Cells to Mitochondrial Pro-Oxidant Insult
title_full_unstemmed SSADH Variants Increase Susceptibility of U87 Cells to Mitochondrial Pro-Oxidant Insult
title_short SSADH Variants Increase Susceptibility of U87 Cells to Mitochondrial Pro-Oxidant Insult
title_sort ssadh variants increase susceptibility of u87 cells to mitochondrial pro-oxidant insult
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7352567/
https://www.ncbi.nlm.nih.gov/pubmed/32575506
http://dx.doi.org/10.3390/ijms21124374
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