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...

Full description

Bibliographic Details
Main Authors: María G Mediavilla, Gisela A Di Venanzio, Edgardo E Guibert, Claudio Tiribelli
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2010-01-01
Series:PLoS ONE
Online Access:http://europepmc.org/articles/PMC2957446?pdf=render
id doaj-c22bddc7ac6046c2b57b73ec0e63001a
record_format Article
spelling 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 AT mariagmediavilla heterologousferredoxinreductaseandflavodoxinprotectcos7cellsfromoxidativestress
AT giselaadivenanzio heterologousferredoxinreductaseandflavodoxinprotectcos7cellsfromoxidativestress
AT edgardoeguibert heterologousferredoxinreductaseandflavodoxinprotectcos7cellsfromoxidativestress
AT claudiotiribelli heterologousferredoxinreductaseandflavodoxinprotectcos7cellsfromoxidativestress
_version_ 1725115484398944256