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.

Menduti, G., Vitaliti, A., Capo, C.r., Lettieri-Barbato, D., Aquilano, K., Malaspina, P., et al. (2020). SSADH variants increase susceptibility of U87 cells to mitochondrial pro-oxidant insult. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, 21(12) [10.3390/ijms21124374].

SSADH variants increase susceptibility of U87 cells to mitochondrial pro-oxidant insult

Menduti, Giovanna
Investigation
;
Capo, Concetta Rosa
Investigation
;
Lettieri-Barbato, Daniele
Investigation
;
Aquilano, Katia
Conceptualization
;
Malaspina, Patrizia
Conceptualization
;
Rossi, Luisa
Conceptualization
2020-06-19

Abstract

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.
19-giu-2020
Pubblicato
Rilevanza internazionale
Articolo
Esperti anonimi
Settore BIO/12 - BIOCHIMICA CLINICA E BIOLOGIA MOLECOLARE CLINICA
Settore BIO/18 - GENETICA
Settore BIO/10 - BIOCHIMICA
English
Con Impact Factor ISI
ALDH5A1
GABA
SSA
SSADH
U87 cells
mitochondria
paraquat
4-HNE
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7352567/pdf/ijms-21-04374.pdf
Menduti, G., Vitaliti, A., Capo, C.r., Lettieri-Barbato, D., Aquilano, K., Malaspina, P., et al. (2020). SSADH variants increase susceptibility of U87 cells to mitochondrial pro-oxidant insult. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, 21(12) [10.3390/ijms21124374].
Menduti, G; Vitaliti, A; Capo, Cr; Lettieri-Barbato, D; Aquilano, K; Malaspina, P; Rossi, L
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2108/252553
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