• Home
  • Search Results
  • Functional interaction of glutathione S-transferase pi and peroxiredoxin 6 in intact cells.

Functional interaction of glutathione S-transferase pi and peroxiredoxin 6 in intact cells.

The international journal of biochemistry & cell biology (2012-11-21)
Suiping Zhou, Yu-Chin Lien, Tea Shuvaeva, Kristine DeBolt, Sheldon I Feinstein, Aron B Fisher
ABSTRACT

Peroxiredoxin 6 (Prdx6) is a 1-Cys member of the peroxiredoxin superfamily that plays an important role in antioxidant defense. Glutathionylation of recombinant Prdx6 mediated by π glutathione S-transferase (GST) is required for reduction of the oxidized Cys and completion of the peroxidatic catalytic cycle in vitro. This study investigated the requirement for πGST in intact cells. Transfection with a plasmid construct expressing πGST into MCF7, a cell line that lacks endogenous πGST, significantly increased phospholipid peroxidase activity as measured in cell lysates and protected intact cells against a peroxidative stress. siRNA knockdown indicated that this increased peroxidase activity was Prdx6 dependent. Interaction between πGST and Prdx6, evaluated by the Duolink Proximity Ligation Assay, was minimal under basal conditions but increased dramatically following treatment of cells with the oxidant, tert-butyl hydroperoxide. Interaction was abolished by mutation of C47, the active site for Prdx6 peroxidase activity. Depletion of cellular GSH by treatment of cells with buthionine sulfoximine had no effect on the interaction of Prdx6 and πGST. These data are consistent with the hypothesis that oxidation of the catalytic cysteine in Prdx6 is required for its interaction with πGST and that the interaction plays an important role in regenerating the peroxidase activity of Prdx6.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Duolink® In Situ Red Starter Kit Mouse/Rabbit
Sigma-Aldrich
Duolink® In Situ Detection Reagents Red
Sigma-Aldrich
Duolink® In Situ PLA® Probe Anti-Mouse MINUS, Affinity purified Donkey anti-Mouse IgG (H+L)
Sigma-Aldrich
Duolink® In Situ PLA® Probe Anti-Rabbit PLUS, Affinity purified Donkey anti-Rabbit IgG (H+L)
Sigma-Aldrich
Duolink® In Situ Orange Starter Kit Mouse/Rabbit
Sigma-Aldrich
Duolink® In Situ PLA® Probe Anti-Mouse PLUS
Sigma-Aldrich
Duolink® In Situ PLA® Probe Anti-Rabbit MINUS
Sigma-Aldrich
Duolink® In Situ Detection Reagents Orange
Sigma-Aldrich
Duolink® In Situ Detection Reagents Green
Sigma-Aldrich
Duolink® In Situ Detection Reagents FarRed
Sigma-Aldrich
Duolink® In Situ Probemaker MINUS
Sigma-Aldrich
Duolink® In Situ Probemaker PLUS
Sigma-Aldrich
Duolink® In Situ Detection Reagents Brightfield
Sigma-Aldrich
Duolink® In Situ PLA® Probe Anti-Goat PLUS
Sigma-Aldrich
Duolink® In Situ PLA® Probe Anti-Goat MINUS, Affinity purified Donkey anti-Goat IgG (H+L)
Sigma-Aldrich
Duolink® In Situ Red Starter Kit Mouse/Goat
Sigma-Aldrich
Duolink® In Situ Red Starter Kit Goat/Rabbit
Sigma-Aldrich
Duolink® In Situ Orange Starter Kit Goat/Rabbit
Sigma-Aldrich
Duolink® In Situ Orange Starter Kit Mouse/Goat