Construction and activity analyses of single functional mouse peroxiredoxin 6 (Prdx6)

Abstract Introduction: Peroxiredoxin 6 (Prdx6) is a bifunctional protein with glutathione peroxidase activity and phospholipase A2 activity. Previous studies have shown a significant positive correlation between the intracellular survival ability of Brucella and Prdx6. Here, the Prdx6 enzyme with a single activity was constructed to facilitate study of the relationship between the single function of Prdx6 and Brucella infection. Material and Methods: The target open reading frame (ORF) DNAs of Prdx6 with a single active centre were prepared using gene splicing by overlap extension PCR (SOE-PCR), and the recombinant eukaryotic expression plasmids inserted by Prdx6 with the single activity centre were constructed and transfected into murine Raw264.7 macrophages. The glutathione peroxidase activity and phospholipase A2 activity of the constructed Prdx6 were examined. Results: The core centres (Ser32 and Cys47) of Prdx6 were successfully mutated by changing the 94th nucleotide from T to G and the 140th nucleotide from G to C in the two enzyme activity cores, respectively. The constructed recombinant plasmids of Prdx6 with the single active centre were transfected into murine macrophages showing the expected single functional enzyme activity, which MJ33 or mercaptosuccinate inhibitors were able to inhibit. Conclusion: The constructed mutants of Prdx6 with the single activity cores will be a benefit to further study of the biological function of Prdx6 with different enzyme activity.

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Ethnic variation in red cell glutathione peroxidase activity

Blood ◽

10.1182/blood.v46.1.103.103 ◽

1975 ◽

Vol 46 (1) ◽

pp. 103-110 ◽

Cited By ~ 43

Author(s):

E Beutler ◽

F Matsumoto

Keyword(s):

Enzyme Activity ◽

Glutathione Peroxidase ◽

Peroxidase Activity ◽

Genetic Model ◽

The United States ◽

European Population ◽

Red Cell ◽

Glutathione Peroxidase Activity ◽

Cultured Fibroblasts ◽

Erythrocyte Enzyme

Abstract Glutathione peroxidase activity was measured in blood and cultured fibroblasts from healthy persons of several different population groups. Individuals of Jewish ancestry and others of Mediterranean origin were found to manifest a decrease of red cell but not of leukocyte or fibroblast enzyme activity. Oriental populations differed in that the scatter in red cell enzyme activity was significantly lower than in Occidental populations. The erythrocyte enzyme of individuals with low activity was found to be less stable to heating than was the enzyme from persons with high activity. As a possible explanation for these data, a provisional genetic model is presented: a low GSH Px allele with a frequency of 0.556 in the Jewish population and of only 0.181 in the United States-Northern European population. Our results suggest that an association between GSH Px deficiency and hemolytic anemia need not represent a cause-and-effect relationship.

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Decreased Glutathione Peroxidase Activity Secondary to Severe Iron Deficiency: A Possible Mechanism Responsible for the Shortened Life Span of the Iron-deficient Red Cell

Blood ◽

10.1182/blood.v43.2.281.281 ◽

1974 ◽

Vol 43 (2) ◽

pp. 281-289 ◽

Cited By ~ 25

Author(s):

Robert Rodvien ◽

Amanda Gillum ◽

Lewis R. Weintraub

Keyword(s):

Enzyme Activity ◽

Iron Deficiency ◽

Glutathione Peroxidase ◽

Peroxidase Activity ◽

Hemoglobin Concentration ◽

Red Cells ◽

Red Cell ◽

Glutathione Peroxidase Activity ◽

Iron Deficient ◽

Red Cell Survival

Abstract Red cell glutathione peroxidase activity decreased to 26% of initial activity following the induction of severe iron deficiency in rabbits. The fall in enzyme activity does not appear to be related to anemia per se, aging of the animals during the course of the experiment, or a generalized decrease in enzyme protein synthesis. With iron repletion, glutathione peroxidase activity did not return to initial values for 10 wk after the hemoglobin concentration had returned to normal. During this period of time, the young red cell population had a higher concentration of GSH-Px activity than the older cells. In contrast, there is no age differential in the concentration of this enzyme in normal red cells, and during the induction of iron deficiency young red cells have decreased enzyme activity as compared to older cells. Reduction in glutathione peroxidase activity may result in increased oxidative damage to the iron-deficient red cell. This, in turn, may be responsible for the previously reported membrane abnormalities and shortened red cell survival of iron-deficient red cells. It remains to be shown if glutathione peroxidase is dependent on iron for its synthesis or if the function of the enzyme is dependent on an iron-containing protein serving as an electron carrier.

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Ethnic variation in red cell glutathione peroxidase activity

Blood ◽

10.1182/blood.v46.1.103.bloodjournal461103 ◽

1975 ◽

Vol 46 (1) ◽

pp. 103-110

Author(s):

E Beutler ◽

F Matsumoto

Keyword(s):

Enzyme Activity ◽

Glutathione Peroxidase ◽

Peroxidase Activity ◽

Genetic Model ◽

The United States ◽

European Population ◽

Red Cell ◽

Glutathione Peroxidase Activity ◽

Cultured Fibroblasts ◽

Erythrocyte Enzyme

Glutathione peroxidase activity was measured in blood and cultured fibroblasts from healthy persons of several different population groups. Individuals of Jewish ancestry and others of Mediterranean origin were found to manifest a decrease of red cell but not of leukocyte or fibroblast enzyme activity. Oriental populations differed in that the scatter in red cell enzyme activity was significantly lower than in Occidental populations. The erythrocyte enzyme of individuals with low activity was found to be less stable to heating than was the enzyme from persons with high activity. As a possible explanation for these data, a provisional genetic model is presented: a low GSH Px allele with a frequency of 0.556 in the Jewish population and of only 0.181 in the United States-Northern European population. Our results suggest that an association between GSH Px deficiency and hemolytic anemia need not represent a cause-and-effect relationship.

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Faculty Opinions recommendation of Interaction of surfactant protein A with peroxiredoxin 6 regulates phospholipase A2 activity.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1050437.502345 ◽

2007 ◽

Author(s):

Maurizio Luisetti

Keyword(s):

Phospholipase A2 ◽

Protein A ◽

Surfactant Protein ◽

Surfactant Protein A ◽

Peroxiredoxin 6 ◽

Phospholipase A2 Activity

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Low Selenium Status in the Elderly Influences Thyroid Hormones

Clinical Science ◽

10.1042/cs0890637 ◽

1995 ◽

Vol 89 (6) ◽

pp. 637-642 ◽

Cited By ~ 64

Author(s):

Oliviero Olivieri ◽

Domenico Girelli ◽

Margherita Azzini ◽

Anna Maria Stanzial ◽

Carla Russo ◽

...

Keyword(s):

Glutathione Peroxidase ◽

Thyroid Hormones ◽

Peroxidase Activity ◽

Controlled Trial ◽

The Elderly ◽

Serum Selenium ◽

Selenium Status ◽

Glutathione Peroxidase Activity ◽

Double Blind ◽

Double Blind Placebo

1. Iodothyronine 5′-deiodinase, which is mainly responsible for peripheral triiodothyronine (T3) production, has recently been demonstrated to be a selenium-containing enzyme. In the elderly, reduced peripheral conversion of thyroxine (T4) to T3 and overt hypothyroidism are frequently observed. 2. We measured serum selenium and erythrocyte glutathione peroxidase (as indices of selenium status), thyroid hormones and thyroid-stimulating hormone in 109 healthy euthyroid subjects (52 women, 57 men), carefully selected to exclude abnormally low thyroid hormone levels induced by acute or chronic diseases or calorie restriction. The subjects were subdivided into three age groups. To avoid conditions of undernutrition or malnutrition, dietary records were obtained for a sample of 24 subjects, randomly selected and representative of the whole population for age and sex. 3. In order to properly assess the influence of selenium status on iodothyronine 5′-deiodinase type I activity, a double-blind placebo-controlled trial was also carried out on 36 elderly subjects, resident at a privately owned nursing home. 4. In the free-living population, a progressive reduction of the T3/T4 ratio (due to increased T4 levels) and of selenium and erythrocyte glutathione peroxidase activity was observed with advancing age. A highly significant linear correlation between T4, T3/T4 and selenium was observed in the population as a whole (for T4, R = −0.312, P < 0.002; for T3/T4 ratio, R = 0.32, P < 0.01) and in older subjects (for T4, R = −0.40, P < 0.05; for T3/T4 ratio, R = 0.54, P < 0.002). 5. The main result of the double-blind placebo-controlled trial was a significant improvement of selenium indices and a decrease in the T4 level in selenium-treated subjects; serum selenium, erythrocyte glutathione peroxidase activity and thyroid hormones did not change in placebo-treated subjects. 6. We concluded that selenium status influences thyroid hormones in the elderly, mainly modulating T4 levels.

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Selenium levels and glutathione peroxidase activity in patients with ataxia-telangiectasia: association with oxidative stress and lipid status biomarkers

Orphanet Journal of Rare Diseases ◽

10.1186/s13023-021-01732-5 ◽

2021 ◽

Vol 16 (1) ◽

Author(s):

Itana Gomes Alves Andrade ◽

Fabíola Isabel Suano-Souza ◽

Fernando Luiz Affonso Fonseca ◽

Carolina Sanchez Aranda Lago ◽

Roseli Oselka Saccardo Sarni

Keyword(s):

Oxidative Stress ◽

Glutathione Peroxidase ◽

Peroxidase Activity ◽

Oxidized Ldl ◽

Reference Value ◽

Glutathione Peroxidase Activity ◽

Oxidative Stress Biomarkers ◽

Apo B ◽

Lipid Status ◽

And Oxidative Stress

Abstract Introduction Ataxia-Telangiectasia (A-T) is a multi-system disorder that may be associated with endocrine changes, oxidative stress in addition to inflammation. Studies suggest that selenium is a trace element related to protection against damage caused by oxidative stress. Objective To describe the plasma levels of selenium and erythrocyte glutathione peroxidase activity in A-T patients and to relate them to oxidative stress and lipid status biomarkers. Methods This is a cross-sectional and controlled study evaluating 22 A-T patients (age median, 12.2 years old) matched by gender and age with 18 healthy controls. We evaluated: nutritional status, food intake, plasma selenium levels, erythrocyte glutathione peroxidase activity, lipid status, inflammation and oxidative stress biomarkers. Results Adequate levels of selenium were observed in 24/36 (66.7%) in this evaluated population. There was no statistically significant difference between the groups in selenium levels [47.6 μg/L (43.2–57.0) vs 54.6 (45.2–62.6) μg/dL, p = 0.242]. Nine of A-T patients (41%) had selenium levels below the reference value. The A-T group presented higher levels of LDL-c, non-HDL-c, oxidized LDL, Apo B, Apo-B/Apo-A-I1, LDL-c/HDL-c ratio, malondialdehyde [3.8 µg/L vs 2.8 µg/L, p = 0.029] and lower Apo-A-I1/HDL-c and glutathione peroxidase activity [7300 U/L vs 8686 U/L, p = 0.005]. Selenium levels were influenced, in both groups, independently, by the concentrations of oxidized LDL, malonaldehyde and non-HDL-c. The oxidized LDL (AUC = 0.849) and ALT (AUC = 0.854) were the variables that showed the greatest discriminatory power between groups. Conclusion In conclusion, we observed the presence of selenium below the reference value in nearly 40% and low GPx activity in A-T patients. There was a significant, inverse and independent association between selenium concentrations and oxidative stress biomarkers. Those data reinforce the importance of assessing the nutritional status of selenium in those patients.

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The distribution, induction and isoenzyme profile of glutathione S-transferase and glutathione peroxidase in isolated rat liver parenchymal, Kupffer and endothelial cells

Biochemical Journal ◽

10.1042/bj2640737 ◽

1989 ◽

Vol 264 (3) ◽

pp. 737-744 ◽

Cited By ~ 18

Author(s):

P Steinberg ◽

H Schramm ◽

L Schladt ◽

L W Robertson ◽

H Thomas ◽

...

Keyword(s):

Endothelial Cells ◽

Glutathione Peroxidase ◽

Rat Liver ◽

Peroxidase Activity ◽

Cell Types ◽

Transferase Activity ◽

Glutathione Peroxidase Activity ◽

Glutathione S Transferase ◽

Liver Parenchymal ◽

Parenchymal Cells

The distribution and inducibility of cytosolic glutathione S-transferase (EC 2.5.1.18) and glutathione peroxidase (EC 1.11.1.19) activities in rat liver parenchymal, Kupffer and endothelial cells were studied. In untreated rats glutathione S-transferase activity with 1-chloro-2,4-dinitrobenzene and 4-hydroxynon-2-trans-enal as substrates was 1.7-2.2-fold higher in parenchymal cells than in Kupffer and endothelial cells, whereas total, selenium-dependent and non-selenium-dependent glutathione peroxidase activities were similar in all three cell types. Glutathione S-transferase isoenzymes in parenchymal and non-parenchymal cells isolated from untreated rats were separated by chromatofocusing in an f.p.l.c. system: all glutathione S-transferase isoenzymes observed in the sinusoidal lining cells were also detected in the parenchymal cells, whereas Kupffer and endothelial cells lacked several glutathione S-transferase isoenzymes present in parenchymal cells. At 5 days after administration of Arocolor 1254 glutathione S-transferase activity was only enhanced in parenchymal cells; furthermore, selenium-dependent glutathione peroxidase activity decreased in parenchymal and non-parenchymal cells. At 13 days after a single injection of Aroclor 1254 a strong induction of glutathione S-transferase had taken place in all three cell types, whereas selenium-dependent glutathione peroxidase activity remained unchanged (endothelial cells) or was depressed (parenchymal and Kupffer cells). Hence these results clearly establish that glutathione S-transferase and glutathione peroxidase are differentially regulated in rat liver parenchymal as well as non-parenchymal cells. The presence of glutathione peroxidase and several glutathione S-transferase isoenzymes capable of detoxifying a variety of compounds in Kupffer and endothelial cells might be crucial to protect the liver from damage by potentially hepatotoxic substances.

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