Activity assay of mammalian 2-cys peroxiredoxins using yeast thioredoxin reductase system

Kim, Ju-A.; Park, Sunjoo; Kim, Kangwha; Rhee, Sue Goo; Kang, Sang Won

doi:10.1016/j.ab.2004.12.008

Cited by 57 publications

(48 citation statements)

References 38 publications

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“…And it is reported that acute exposure induces anatomical lesions at the proximal tubular cells and lipid peroxidation in human kidney (Hose et al, 2016).Chromium reduced intermediates were thought to reacted with hydrogen peroxide to form hydroxyl radicals (Bucher, 2007), with subsequent alterations in proteins, DNA, and phospho-lipids leading to disturbing cellular functions and its integrity (Bucher, 2007). So that, rats were intoxicated with K 2 Cr 2 O 7 with subsequent increase in the serum creatinine, urea, uric acid, proteins and sodium levels and severe alteration in the histopathological examination in comparison with normal control group, through the elevation of reactive oxygen species (ROS) that induces tissues damage such as liver, pancreas, cerebellum and kidney (Kim et al, 2005;Fatima et al, 2005). ROS generated by this process can bring on injury to cellular proteins, lipids, and DNA leading to oxidative stress (Li et al, 2008;Mehany et al, 2013).…”

Section: Discussionmentioning

confidence: 99%

Amelioration of the nephrotoxic effect of potassium dichromate by whey protein and/or nigella sativa oil in male albino rats

Bashandy¹,

Amin²,

Morsy³

et al. 2016

J App Pharm Sci

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Even though whey protein complex derived from milk, is being touted as a functional food with a number of health benefits, so far its full mechanistic effect has not been deeply explicated, which is the target of this study. For this reason, we observed the 2 months protection influence of whey protein (100 & 200 mg/kg, p.ob.wt.) and/or oil of Nigella sativa (5ml/kg, p.ob.wt.) against intoxicated rats with single dose of potassium dichromate (30mg/Kg, I.P) at the end of protection period. On the biochemical level, whey protein and/or oil of Nigella sativa were changed the potassium dichromate renal toxicity consequences, where it showed a significant improvement in the creatinine, urea, protein, uric acid and sodium levels. While in the tissue level (Kidney), it revealed a significant enhancement in reduced glutathione (GSH), catalase and superoxide dismutase (SOD) enzymes as compared to nephrotoxic group. Moreover, whey protein and/or oil of Nigella sativa reduced the elevation of malondialdehyde (MDA) and inflammatory mediators like tumor necrotic factor-alpha (TNF-α) and nitric oxide (NO) induced by potassium dichromate and alleviation the histopathological changes caused by the intoxication. These results give a new vision into the hopeful mechanisms that modulated numerous factors induced renal injury.

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Section: Discussionmentioning

confidence: 99%

Amelioration of the nephrotoxic effect of potassium dichromate by whey protein and/or nigella sativa oil in male albino rats

Bashandy¹,

Amin²,

Morsy³

et al. 2016

J App Pharm Sci

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“…The fractions containing Prx1 or Prx1C52S were pooled and stored at À80jC. Yeast thioredoxin and thioredoxin reductase proteins were prepared following the previously described procedure (31).…”

Section: Methodsmentioning

confidence: 99%

Prx1 Suppresses Radiation-Induced c-Jun NH2-Terminal Kinase Signaling in Lung Cancer Cells through Interaction with the Glutathione S-Transferase Pi/c-Jun NH2-Terminal Kinase Complex

Kim¹,

Lee²,

et al. 2006

Cancer Research

130

119

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Radiotherapy is one of the major treatment modalities for lung cancer. Cell killing by ionizing radiation is mediated primarily through the reactive oxygen species (ROS) and ROSdriven oxidative stress. Prx1, a peroxiredoxin family member, was shown to be frequently elevated in lung cancer cells and tissues. Although the antioxidant function of Prx1 is expected to affect the radiotherapy response of lung cancer, the physiologic significance of its peroxidase activity in irradiated cells is unclear because the catalytic Cys 52 is easily inactivated by ROS due to its overoxidation to sulfinic or sulfonic acid. In this study, we investigated the role of Prx1 in radiation sensitivity of human lung cancer cells, with special emphasis on the redox status of the catalytic Cys 52 . We found that overexpression of Prx1 enhances the clonogenic survival of irradiated cells and suppresses ionizing radiation-induced c-Jun NH 2 -terminal kinase (JNK) activation and apoptosis. The peroxidase activity of Prx1, however, is not essential for inhibiting JNK activation. The latter effect is mediated through its association with the glutathione S-transferase pi (GSTpi)-JNK complex, thereby preventing JNK release from the complex. Reduced JNK activation is observed when the peroxidase activity of Prx1 is compromised by Cys 52 overoxidation or in the presence of the Cys 52 to Ser 52 mutant (Prx1C52S) lacking peroxidase activity. We show that both Prx1 and Prx1C52S interact with the GSTpi-JNK complex and suppress the release of JNK from the complex. Our study provides new insight into the antiapoptotic function of Prx1 in modulating radiosensitivity and provides the impetus to monitor the influence of Prx1 levels in the management of lung cancer. (Cancer Res 2006; 66(14): 7136-42)

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“…To characterize this activity and to demonstrate that PrxIV can act as a peroxidase, we prepared recombinant protein by expression in E. coli. Using a standard peroxide-turnover assay (Kim et al, 2005), we showed that human PrxIV exhibits peroxidase activity in vitro ( Fig. 2A).…”

Section: Prxiv Has Peroxidase Activity Both In Isolation and Within Cmentioning

confidence: 99%

“…His-tags were cleaved with 10 U thrombin protease (GE Healthcare, Amersham, UK) per mg purified protein for 16 hours at 4° C and PrxIV decamers then separated to homogeneity using Superdex 200 TM column chromatography (GE Healthcare). PrxIV activity assays and step-wise reduction by thioredoxin were performed using previously established reaction conditions (Kim et al, 2005). Final concentrations were 4.5 M PrxIV, 3 M thioredoxin, 1.5 M thioredoxin reductase, 200 M NADPH and 100 M hydrogen peroxide.…”

Section: In Vitro Analysesmentioning

confidence: 99%

Peroxiredoxin IV protects cells from oxidative stress by removing H2O2 produced during disulphide formation

Tavender

Bulleid

2010

Journal of Cell Science

152

155

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SummaryDisulphide formation within the endoplasmic reticulum (ER) requires the activity of the ER oxidase Ero1, and as a consequence, generates hydrogen peroxide. The production of hydrogen peroxide is thought to lead to oxidative stress that ultimately results in apoptosis. Here, we show that mammalian peroxiredoxin IV (PrxIV) metabolises hydrogen peroxide produced by Ero1. We demonstrate that the presence of PrxIV within the ER provides a cytoprotective effect against stresses known to enhance Ero1 activity and perturb ER redox balance. Increased Ero1 activity and production of hydrogen peroxide led to preferential hyperoxidation of PrxIV relative to peroxiredoxins in other cellular compartments. The hyperoxidation was increased by the upregulation of Ero1 and by the expression of a hyperactive Ero1. These findings provide the first evidence for an enzymatic mechanism that facilitates peroxide removal from the ER, and show that the oxidation status of PrxIV acts as a marker for ER oxidative stress.

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Activity assay of mammalian 2-cys peroxiredoxins using yeast thioredoxin reductase system

Cited by 57 publications

References 38 publications

Amelioration of the nephrotoxic effect of potassium dichromate by whey protein and/or nigella sativa oil in male albino rats

Amelioration of the nephrotoxic effect of potassium dichromate by whey protein and/or nigella sativa oil in male albino rats

Prx1 Suppresses Radiation-Induced c-Jun NH2-Terminal Kinase Signaling in Lung Cancer Cells through Interaction with the Glutathione S-Transferase Pi/c-Jun NH2-Terminal Kinase Complex

Peroxiredoxin IV protects cells from oxidative stress by removing H2O2 produced during disulphide formation

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