Heterologous ferredoxin reductase and flavodoxin protect Cos-7 cells from oxidative stress.
BACKGROUND: Ferredoxin-NADP(H) reductase (FNR) from Pisum sativum and Flavodoxin (Fld) from Anabaena PCC 7119 have been reported to protect a variety of cells and organisms from oxidative insults. In this work, these two proteins were expressed in mitochondria of Cos-7 cells and tested for their eff...
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doaj-c22bddc7ac6046c2b57b73ec0e63001a2020-11-25T01:25:03ZengPublic Library of Science (PLoS)PLoS ONE1932-62032010-01-01510e1350110.1371/journal.pone.0013501Heterologous ferredoxin reductase and flavodoxin protect Cos-7 cells from oxidative stress.María G MediavillaGisela A Di VenanzioEdgardo E GuibertClaudio TiribelliBACKGROUND: Ferredoxin-NADP(H) reductase (FNR) from Pisum sativum and Flavodoxin (Fld) from Anabaena PCC 7119 have been reported to protect a variety of cells and organisms from oxidative insults. In this work, these two proteins were expressed in mitochondria of Cos-7 cells and tested for their efficacy to protect these cells from oxidative stress in vitro. PRINCIPAL FINDINGS: Cos-7/pFNR and Cos-7/pFld cell lines expressing FNR and Fld, respectively, showed a significantly higher resistance to 24 h exposure to 300-600 µM hydrogen peroxide measured by LDH retention, MTT reduction, malondialdehyde (MDA) levels and lipid peroxide (LPO; FOX assay) levels. However, FNR and Fld did not exhibit any protection at shorter incubation times (2 h and 4 h) to 4 mM hydrogen peroxide or to a 48 h exposure to 300 µM methyl viologen. We found enhanced methyl viologen damage exerted by FNR that may be due to depletion of NADPH pools through NADPH-MV diaphorase activity as previously observed for other overexpressed enzymes. SIGNIFICANCE: The results presented are a first report of antioxidant function of these heterologous enzymes of vegetal and cyanobacterial origin in mammalian cells.http://europepmc.org/articles/PMC2957446?pdf=render |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
María G Mediavilla Gisela A Di Venanzio Edgardo E Guibert Claudio Tiribelli |
spellingShingle |
María G Mediavilla Gisela A Di Venanzio Edgardo E Guibert Claudio Tiribelli Heterologous ferredoxin reductase and flavodoxin protect Cos-7 cells from oxidative stress. PLoS ONE |
author_facet |
María G Mediavilla Gisela A Di Venanzio Edgardo E Guibert Claudio Tiribelli |
author_sort |
María G Mediavilla |
title |
Heterologous ferredoxin reductase and flavodoxin protect Cos-7 cells from oxidative stress. |
title_short |
Heterologous ferredoxin reductase and flavodoxin protect Cos-7 cells from oxidative stress. |
title_full |
Heterologous ferredoxin reductase and flavodoxin protect Cos-7 cells from oxidative stress. |
title_fullStr |
Heterologous ferredoxin reductase and flavodoxin protect Cos-7 cells from oxidative stress. |
title_full_unstemmed |
Heterologous ferredoxin reductase and flavodoxin protect Cos-7 cells from oxidative stress. |
title_sort |
heterologous ferredoxin reductase and flavodoxin protect cos-7 cells from oxidative stress. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS ONE |
issn |
1932-6203 |
publishDate |
2010-01-01 |
description |
BACKGROUND: Ferredoxin-NADP(H) reductase (FNR) from Pisum sativum and Flavodoxin (Fld) from Anabaena PCC 7119 have been reported to protect a variety of cells and organisms from oxidative insults. In this work, these two proteins were expressed in mitochondria of Cos-7 cells and tested for their efficacy to protect these cells from oxidative stress in vitro. PRINCIPAL FINDINGS: Cos-7/pFNR and Cos-7/pFld cell lines expressing FNR and Fld, respectively, showed a significantly higher resistance to 24 h exposure to 300-600 µM hydrogen peroxide measured by LDH retention, MTT reduction, malondialdehyde (MDA) levels and lipid peroxide (LPO; FOX assay) levels. However, FNR and Fld did not exhibit any protection at shorter incubation times (2 h and 4 h) to 4 mM hydrogen peroxide or to a 48 h exposure to 300 µM methyl viologen. We found enhanced methyl viologen damage exerted by FNR that may be due to depletion of NADPH pools through NADPH-MV diaphorase activity as previously observed for other overexpressed enzymes. SIGNIFICANCE: The results presented are a first report of antioxidant function of these heterologous enzymes of vegetal and cyanobacterial origin in mammalian cells. |
url |
http://europepmc.org/articles/PMC2957446?pdf=render |
work_keys_str_mv |
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