scholarly journals Glutathione and ascorbic acid in spinach (Spinacia oleracea) chloroplasts. The effect of hydrogen peroxide and of Paraquat

1983 ◽  
Vol 210 (3) ◽  
pp. 899-903 ◽  
Author(s):  
M Y Law ◽  
S A Charles ◽  
B Halliwell
Keyword(s):  
Hydrogen Peroxide ◽  
Ascorbic Acid ◽  
Spinacia Oleracea ◽  
Reduced Glutathione ◽  
Oxidized Glutathione ◽  
Fructose Bisphosphatase ◽  
Physiological Conditions ◽  
Spinach Chloroplasts ◽  
Apparent Loss

The stroma of spinach chloroplasts contains ascorbic acid and glutathione at millimolar concentrations. [Reduced glutathione]/[oxidized glutathione] and [ascorbate]/[dehydroascorbate] ratios are high under both light and dark conditions and no evidence for a role of oxidized glutathione or dehydroascorbate in the dark-deactivation of fructose bisphosphatase could be obtained. Addition of H2O2 to chloroplasts in the dark decreases the above ratios, an effect that is reversed on illumination. Addition of Paraquat to illuminated chloroplasts caused a rapid oxidation of reduced glutathione and ascorbate, and apparent loss of dehydroascorbate. Paraquat rapidly inactivated fructose bisphosphatase activity, as assayed under physiological conditions.

THE ROLE OF PLANT PHENOLIC COMPOUNDS IN THE OXIDATION OF REDUCED DIPHOSPHOPYRIDINE NUCLEOTIDE BY PEROXIDASE

1961 ◽  
Vol 39 (7) ◽  
pp. 1113-1124 ◽  
Author(s):  
O. L. Gamborg ◽  
L. R. Wetter ◽  
A. C. Neish
Keyword(s):  
Hydrogen Peroxide ◽  
Ascorbic Acid ◽  
Indoleacetic Acid ◽  
Enzyme System ◽  
Reduced Glutathione ◽  
Maximum Activity ◽  
Coumaric Acid ◽  
Horse Radish Peroxidase ◽  
Competitive Inhibitors

Horse-radish peroxidase and dialyzed extracts from pea epicotyls and from spruce shoots oxidized reduced diphosphopyridine nucleotide in the presence of p-coumaric acid. Maximum activity was obtained when hydrogen peroxide, Mn+2, p-coumaric acid, and enzyme were present. p-Hydroxyphenylpropionic acid, p-hydroxyphenylpyruvic acid, tyrosol, or resorcinol could replace p-coumaric acid as the phenolic activator but they were less efficient. Ferulic and sinapic acids were competitive inhibitors while chlorogenic acid, caffeic acid, and hydroquinone were non-competitive inhibitors of the reaction. The oxidation of the reduced coenzyme was inhibited by citrate and pyrophosphate, enhanced by versene, and delayed by ascorbic acid, cysteine, and reduced glutathione. The results indicate that the peroxidase system may be identical with the enzyme system which oxidizes indoleacetic acid.

An ESR study of the peroxidase reaction catalyzed by human methaemoglobin and methaemoglobin-haptoglobin complex

1991 ◽  
Vol 56 (4) ◽  
pp. 923-932
Author(s):  
Jana Stejskalová ◽  
Pavel Stopka ◽  
Zdeněk Pavlíček
Keyword(s):  
Hydrogen Peroxide ◽  
Ascorbic Acid ◽  
Amino Acid ◽  
Peroxidase Activity ◽  
Amino Acid Analysis ◽  
Superoxide Radical ◽  
Esr Spectra ◽  
The Other ◽  
Delocalized Electron

The ESR spectra of peroxidase systems of methaemoglobin-ascorbic acid-hydrogen peroxide and methaemoglobin-haptoglobin complex-ascorbic acid-hydrogen peroxide have been measured in the acetate buffer of pH 4.5. For the system with methaemoglobin an asymmetrical signal with g ~ 2 has been observed which is interpreted as the perpendicular region of anisotropic spectrum of superoxide radical. On the other hand, for the system with methaemoglobin-haptoglobin complex the observed signal with g ~ 2 is symmetrical and is interpreted as a signal of delocalized electron. After realization of three repeatedly induced peroxidase processes the ESR signal of the perpendicular part of anisotropic spectrum of superoxide radical is distinctly diminished, whereas the signal of delocalized electron remains practically unchanged. An amino acid analysis of methaemoglobin along with results of the ESR measurements make it possible to derive a hypothesis about the role of haptoglobin in increasing of the peroxidase activity of methaemoglobin.

The formation of choleglobin and the role of catalase in the erythrocyte

1949 ◽  
Vol 136 (884) ◽  
pp. 435-448 ◽  
Keyword(s):  
Hydrogen Peroxide ◽  
Ascorbic Acid ◽  
Amino Acid ◽  
Catalase Activity ◽  
Acid Oxidase ◽  
Small Decrease ◽  
Amino Acid Oxidase ◽  
Oxidase System ◽  
Inverse Proportion

In haemolysates of non-nucleated erythrocytes there is an inverse proportion between catalase activity and rate of choleglobin formation on addition of ascorbic acid. In the intact erythrocytes catalase protects haemoglobin against oxidation and further destruction by the hydrogen peroxide generated by the D-amino-acid oxidase system or by physiological concentrations of ascorbic acid and glutathione. Acid destromatization of haemolyzed horse erythrocytes causes a small decrease in the catalase activity and an increased rate of inactivation of the remaining catalase by ascorbic acid. The liberation of copper from haemocuprein is quantitatively insufficient to explain the decreased stability of the catalase. Exposing duck oxyhaemoglobin, but not reduced haemoglobin, to a pH of 5⋅5 to 5⋅8, causes an alteration which is apparent from the increase of the rate of choleglobin formation. The mechanism of this alteration is discussed. It partly explains the 'stroma effect', at least in duck erythrocytes. In addition, in the latter, there is a true stroma effect. Choleglobin formation in the presence of ascorbic acid is accelerated by a variety of substances. Some of these perturb haemoglobin, while others increase the formation of hydrogen peroxide from ascorbic acid. The implications of our findings on the mechanism of choleglobin formation and on the role of catalase in the erythrocyte are discussed.

scholarly journals Action of calcium ions on spinach (Spinacia oleracea) chloroplast fructose bisphosphatase and other enzymes of the Calvin cycle

1980 ◽  
Vol 188 (3) ◽  
pp. 775-779 ◽  
Author(s):  
S A Charles ◽  
B Halliwell
Keyword(s):  
Calcium Ions ◽  
Spinacia Oleracea ◽  
Calvin Cycle ◽  
Complete Inhibition ◽  
Fructose Bisphosphatase ◽  
Spinach Chloroplasts ◽  
Fructose 1,6 Bisphosphate ◽  
Sedoheptulose Bisphosphatase

Thiol-treated spinach (Spinacia oleracea) chloroplast fructose bisphosphatase is powerfully inhibited by Ca2+ non-competitively with respect to its substrate, fructose 1,6-bisphosphate. 500 microM-Ca2+ causes virtually complete inhibition and the Ki is 40 microM. Severe inhibition of sedoheptulose bisphosphatase is also caused by Ca2+. A role for Ca2+ in regulation of the Calvin cycle in spinach chloroplasts is proposed.

Erythrocyte defenses against hydrogen peroxide: the role of ascorbic acid

1998 ◽  
Vol 1380 (3) ◽  
pp. 389-395 ◽  
Author(s):  
Shalu Mendiratta ◽  
Zhi-chao Qu ◽  
James M May
Keyword(s):  
Hydrogen Peroxide ◽  
Ascorbic Acid

scholarly journals Reduced Glutathione and Platelet Functions

1977 ◽  
Author(s):  
S. Matsuda ◽  
Y. Ikeda ◽  
M. Kikuchi ◽  
Y. Ando ◽  
M. Aoki ◽  
...  
Keyword(s):  
Platelet Aggregation ◽  
Reduced Glutathione ◽  
Oxidized Glutathione ◽  
Clot Retraction ◽  
Inhibition Of Platelet Aggregation ◽  
Marked Inhibition ◽  
Coomassie Blue ◽  
Specific Agent ◽  
Platelet Functions

The role of reduced glutathione(GSH) on platelet functions was investigated utilizing diamide[(CH3)2NCON=NCON(CH3)2], which was shown to be a rather specific agent for the intracellular oxidation of GSH to oxidized glutathione(GSSG). Human normal citrated PRP was incubated with various concentration of diamide for 30 min at 22°C and platelet aggregation studies were performed using ADP, epinephrine, or collagen as aggregating agents. Clot retraction was examined in PRP treated with diamide by adding 100mM CaCl2 and 1.0u/ml thrombin. Simultaneously, GSH in platelets were measured by Beutler’s method. The maximum aggregability of platelets treated with 25μM diamide was significantly decreased with 2.5μM ADP, 5μM epinephrine, or 64μg/ml collagen as aggregating agent. When platelets were incubated with 50μM diamide, no secondary aggregation with 2.5μM ADP or 5μM epinephrine was observed. At a concentration of 100μM, diamide completely abolished primary aggregation with 2.5μM ADP. Clot retraction was grossly inhibited by diamide at concentrations above 50μM. GSH levels of platelets which were incubated with 100μM and 1mM diamide at 22°C for 30 min was 11.41×l0-9and 1.16×10-9moles/109 plts, respectively, while that without diamide treatment was 15.28×l0-9moles/109 plts. Decrease of GSH levels seemed to be dependent on diamide concentration. Membranes were isolated from diamide treated platelets according to the method of Jamieson et al, and were electrophoreses on SDS Polyacrylamide gels. There was no difference in Coomassie blue or PAS stained bands between control and diamide treated platelets. Marked inhibition of platelet aggregation and clot retraction accompanied by intracellular oxidation of GSH to GSSG may imply an important role of GSH on platelet aggregation and clot retraction.

scholarly journals Oxidative stress and low molecular weight antioxidants in triticale shoots under chloride salinization

2020 ◽  
Vol 62 (6) ◽  
pp. 125-130
Author(s):  
Viktor V. Ivanishchev ◽  
Keyword(s):  
Oxidative Stress ◽  
Hydrogen Peroxide ◽  
Molecular Weight ◽  
Ascorbic Acid ◽  
Correlation Coefficients ◽  
Low Molecular Weight ◽  
Stress Indicators ◽  
Sodium Chloride Stress ◽  
Oxidative Stress Indicators

We studied the alterations in oxidative stress indicators (hydrogen peroxide, superoxide radical, lipid peroxidation – LPO) and the alterations in the content of low molecular weight metabolites (ascorbic acid, glutathione, proline, chlorophyll and carotenoids) in the shoots of triticale (Triticosecale) under short-term (0-96 h) sodium chloride stress (120 mM) with statistical methods: principal component analysis (PCA) and cluster analysis. An analysis of the alterations in oxidative stress indicators allowed us to calculate the correlation coefficients for the pairs: peroxide – superoxide (0.52), peroxide – LPO (0.62), superoxide – LPO (0.23). The inclusion in the analysis of data on alterations in the content of low molecular weight antioxidants showed that the PCA method forms three main groups for all the studied characteristics: (1) LPO and hydrogen peroxide, (2) chlorophyll and carotenoids, (3) glutathione and ascorbate. The correlation coefficients were calculated for pairs: ascorbate – glutathione (0.71), ascorbate – proline (0.81), glutathione – proline (0.28). Such a value of the coefficient of the first pair suggests that ascorbic acid also performs numerous other functions, in addition to participating in the ascorbate-glutathione cycle. The high correlation between ascorbate and proline can be explained by the similar nature of alterations in their content in triticale shoots under conditions of short-term sodium chloride stress. Negative correlation coefficients in pairs of hydrogen peroxide – chlorophyll (-0.73), peroxide – carotenoids (-0.75), ascorbic acid – LPO (-0.70), LPO – proline (-0.69) give reason to talk about the possible protective role of photosynthesis pigments from accumulating hydrogen peroxide, as well as the potential role of ascorbic acid and proline in protecting membranes from lipid peroxidation. The application of the cluster analysis method showed that first and second order clusters between ascorbate, proline and glutathione reflect their known antioxidant role. The results obtained may also indicate that pigments have a much lower protective function.

VARIATION IN GLUTATHIONE AND ASCORBIC ACID CONTENT AMONG SELECTED CULTIVARS OF Phaseolus vulgaris PRIOR TO AND AFTER EXPOSURE TO OZONE

1983 ◽  
Vol 63 (3) ◽  
pp. 733-737 ◽  
Author(s):  
ASAF GURI
Keyword(s):  
Ascorbic Acid ◽  
Phaseolus Vulgaris ◽  
Glutathione Reductase ◽  
Acid Content ◽  
Specific Activity ◽  
Reduced Glutathione ◽  
Ascorbic Acid Content ◽  
Ozone Exposure ◽  
Reduced Form ◽  
Oxidized Glutathione

Measurements of reduced glutathione (GSH) and ascorbic acid (AA) concentrations in the primary leaves of four bean cultivars, two ozone-sensitive (S) and two ozone-insensitive (I), have revealed that GSH concentrations were not significantly different in all four cultivars prior to the exposure to ozone (0.28–0.32 ppm for 8 h); however, after ozone exposure GSH concentrations were significantly lower in the two sensitive cultivars, PHR and 0669, while in the two insensitive tolerants, FH and Nep-2, the drop in GSH concentrations was slight. The assay of glutathione reductase (GR), the enzyme which catalyzed the reduction of oxidized glutathione (GSSG) to its reduced form, namely GSH, indicated that its specific activity in the two insensitive cultivars was almost twice as high as in the two sensitive cultivars. The drop in AA concentration after 8 h of fumigation was moderate (although significant) in all four cultivars, whereas the concentrations of AA in all four cultivars prior to ozone fumigation were not significantly different. From the data it is suggested that differences in GR-specific activity, probably due to different isozymes, could be the reason for insensitivity or sensitivity to ozone among bean cultivars.Key words: Ozone, Phaseolus, glutathione, ascorbic acid, glutathione reductase

scholarly journals Antioxidant System and Biomolecules Alteration in Pisum sativum under Heavy Metal Stress and Possible Alleviation by 5-Aminolevulinic Acid

Molecules ◽  
2019 ◽  
Vol 24 (22) ◽  
pp. 4194 ◽  
Author(s):  
Yasser El-Amier ◽  
Khalid Elhindi ◽  
Salah El-Hendawy ◽  
Sarah Al-Rashed ◽  
Ahmed Abd-ElGawad
Keyword(s):  
Heavy Metals ◽  
Hydrogen Peroxide ◽  
Lipid Peroxidation ◽  
Heavy Metal ◽  
Antioxidant Enzymes ◽  
Pisum Sativum ◽  
Aminolevulinic Acid ◽  
Reduced Glutathione ◽  
Heavy Metal Stress ◽  
Oxidized Glutathione

Environmental pollution is the most serious problem that affects crop productivity worldwide. Pisum sativum is a leguminous plant that is cultivated on a large scale in the Nile Delta of Egypt as a winter crop, and many of the cultivated fields irrigated with drainage water that contained many pollutants including heavy metals. The present research aimed to investigate the impact of Cd and Ni on the biochemical and physiological processes in P. sativum and evaluate the potential alleviation of their toxicity by 5-aminolevulinic acid (ALA). Seedlings of P. sativum were grown in Hoagland solution treated with CdCl2 or NiCl2 for 72 h in the growth chamber. Hydrogen peroxide, lipid peroxidation, protein carbonylation, reduced glutathione, oxidized glutathione, proline, phenolics, antioxidant enzymes, as well as Cd and Ni concentrations were measured at 0, 12, 24, 36, 48, 72 h. An experiment of alleviation was conducted where ALA was added to the growth solution at a concentration of 200 µM coupled with 100 µM of either CdCl2 or NiCl2. Hydrogen peroxide, lipid peroxidation, protein carbonylation, reduced glutathione, oxidized glutathione, proline, and phenolics were induced due to the toxicity of Cd and Ni. The activities of antioxidant enzymes [NADH-oxidase (EC: 1.6.3.1), ascorbate peroxidase (EC: 1.11.1.11), glutathione reductase (EC: 1.6.4.2), superoxide dismutase (EC: 1.15.1.1), and catalase (EC: 1.11.1.6)] were induced under the treatments of both metals. On the other hand, the soluble protein decreased gradually depending upon the time of exposure to the heavy metals. The concentration of Cd and Ni in the leaves treated plants increased in time of exposure dependent manner, while their contents remained within the acceptable limits. The addition of ALA decreased the oxidative stress in treated P. sativum plants. The results revealed the significance of using ALA in the cultivation of P. sativum might improve its tolerance against heavy metal stress.

scholarly journals Antioxidant Responses in Harvested Leaves of Two Cultivars of Spinach Differing in Senescence Rates

2001 ◽  
Vol 126 (5) ◽  
pp. 611-617 ◽  
Author(s):  
D. Mark Hodges ◽  
Charles F. Forney ◽  
Wendy V. Wismer
Keyword(s):  
Hydrogen Peroxide ◽  
Lipid Peroxidation ◽  
Spinacia Oleracea ◽  
Reduced Glutathione ◽  
Total Glutathione ◽  
Antioxidant Responses ◽  
Hydrogen Peroxide Accumulation ◽  
Growth Chambers ◽  
Field Plots ◽  
Postharvest Senescence

The objective of this study was to assess responses of certain antioxidants in harvested leaves of selected cultivars of spinach (Spinacia oleracea L.) differing in postharvest senescence rates in order to explore the significance of these antioxidants in postharvest senescence regulation and dynamics. Ten cultivars were grown in both field plots and laboratory growth chambers, harvested at maturity, and their leaves detached and stored at 10 °C in the dark. Following postharvest analysis, two cultivars were identified consistently as having relatively high (`Spokane F1') and low (`BJ 412 Sponsor') postharvest senescence rates. These two cultivars were then grown in a growth chamber for 45 days and their leaves detached and stored as above. At the point of harvest (day 0) and on days 4, 8, 12, 16, and 20, samples were analyzed for activities of ascorbate peroxidase (ASPX; EC 1.11.1.11), catalase (CAT; EC 1.11.1.6), and superoxide dismutase (SOD; EC 1.15.1.1), and (ii) concentrations of malondialdehyde (MDA, an indicator of lipid peroxidation), total ascorbate, reduced ascorbate (AsA), oxidized ascorbate (DAsA), total glutathione, reduced glutathione (GSH), and oxidized glutathione (GSSG). Although MDA accumulated in leaves of both cultivars concomitant with time after detachment, levels became significantly higher in `Spokane F1'. It is argued that declining activities of ASPX and levels of ascorbate and increasing activities of SOD manifested in accumulation of hydrogen peroxide in `Spokane F1', leading to a greater potential for lipid peroxidation in this cultivar than for `BJ 412 Sponsor'. SOD activities and glutathione levels may have increased as a result of elevated oxidative stress in `Spokane F1'. Increased hydrogen peroxide accumulation in `Spokane F1' relative to `BJ 412 Sponsor' may have contributed to an increased rate of senescence in the harvested leaves of this cultivar.

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