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

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.

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.


1962 ◽  
Vol 40 (1) ◽  
pp. 459-469 ◽  
Author(s):  
P. H. Jellinck ◽  
Louise Irwin

Aerobic incubation of estrone-16-C14with peroxidase in the presence of serum albumin and other proteins resulted in the formation of water-soluble, ether-insoluble metabolites in high percentage yields. Similar products were formed when protein was replaced by cysteine or tryptophan but none of the other amino acids tested had any effect. The evidence points to an initial generation of hydrogen peroxide from these nitrogenous compounds by the enzyme acting as an aerobic oxidase, and the subsequent peroxidation of estrone to highly reactive products. These then combine with the protein or amino acid or else undergo alternative reactions. A strong chemical bond is formed with albumin and attempts to release the estrone metabolites from it were unsuccessful. Uterine homogenates from estrogen-treated rats showing high DPNH oxidase activity contained no "peroxidase" as measured by the formation of water-soluble products from estrone in the presence of protein.


1999 ◽  
Vol 26 (4) ◽  
pp. 387 ◽  
Author(s):  
Francisco J. Pérez ◽  
Verónica Morales

Soluble peroxidase activity from pedicels of seedless table grape cv. Sultana was highly stimulated by post-bloom applications of gibberellic acid (GA3) to vines. The increase in peroxidase activity was mainly due to the induction of a basic peroxidase isoenzyme (pI > 9; BPrx-HpI). The activity of two other peroxidase isoenzymes of pI 6.5 and 3.2 was not altered by the hormone treatment. BPrx-HpI was induced by GA3 in pedicels and rachis but not in berries, although in berries peroxidase activity was also stimulated by post-bloom GA3 applications. BPrx-HpI oxidised guaiacol and ortho-phenylenediamine (o-PDA), while the others peroxidases found in the pedicel and in the berry oxidised only o-PDA. Hence, BPrx-HpI was characterised as a guaiacol-peroxidase showing no activity towards ascorbic acid (ASC). The possible role of BPrx-HpI in pedicel lignification and berry-drop caused by GA3 applications to cv. Sultana vines is discussed.


1981 ◽  
Vol 61 (4) ◽  
pp. 483-486 ◽  
Author(s):  
D. R. Blake ◽  
N. D. Hall ◽  
D. A. Treby ◽  
B. Halliwell ◽  
J. M. C. Gutteridge

1. On exposure of synovial fluid to superoxide and hydrogen peroxide, generated enzymically or by activated polymorphonuclear leucocytes, hyaluronic acid is depolymerized and the fluid loses its lubricating properties. The ability of synovial fluid from rheumatoid patients to scavenge superoxide and hydrogen peroxide was therefore examined. 2. Synovial fluid from a range of rheumatoid patients contained no superoxide dismutase activity, insufficient caeruloplasmin to scavenge any superoxide radical and little, if any, catalase activity. 3. Total ascorbate (reduced ascorbate + dehydroascorbate) concentrations in the plasma and synovial fluid of rheumatoid patients were similar in each case. The values are at the low end of the normal range. 4. These results are discussed in relation to the role of oxygen radicals in inflammatory joint disease.


2020 ◽  
Author(s):  
Vincent Odhiambo Nyasembe ◽  
David Poumo Tchouassi ◽  
Martha Njeri Muturi ◽  
Christian W. W. Pirk ◽  
Catherine L. Sole ◽  
...  

Abstract Background: In a recent study using DNA barcoding, we identified the plants fed upon by four Afro-tropical mosquito species that vector dengue, malaria, and Rift Valley fever. Herein, we have expanded on this study by investigating the role of three of the plants Pithecellobium dulce (Fabaceae), Leonotis nepetifolia (Lamiaceae), and Opuntia ficus-indica (Cactaceae) on survival, fecundity, and egg viability of the dengue vector Aedes aegypti. Methods: We tested these effects using females that received a) an initial three rations of bloodmeal, and b) received no bloodmeal at all. Two controls were included; age-matched females fed on glucose solution with or without initial bloodmeal, and those fed exclusively on bloodmeal. Data was collected daily over a 30-day period. The amino acid content of Ae. aegypti guts and the amino acid content of their respective diets was detected by coupled liquid chromatography – mass spectrometry.Results: Females fed on P. dulce and exclusive bloodmeal had a shorter survival than those fed on glucose. On the other hand, females fed on L. nepetifolia survived longer than those fed exclusively on bloodmeal, whereas those fed on O. ficus-indica had the shortest survival time. With initial bloodmeal, females fed on L. nepetifolia laid 1.6-fold more eggs while those fed on the other diets laid fewer eggs, compared to those fed exclusively on bloodmeal. Hatching rates of the eggs laid varied with the diet. Mass spectroscopic analysis of gut contents of mosquitoes exposed to the different diets showed qualitative and quantitative differences in their amino acid levels. Conclusion: Our findings highlight the central role of plant nutrients in the reproductive fitness of dengue vectors which may impact their disease transmission potential.


2020 ◽  
Author(s):  
Vincent Odhiambo Nyasembe ◽  
David P. Tchouassi ◽  
Martha N. Muturi ◽  
Christian W. W. Pirk ◽  
Catherine L. Sole ◽  
...  

Abstract Background: In a recent study using DNA barcoding, we identified the plants fed upon by four Afro-tropical mosquito species that vector dengue, malaria, and Rift Valley fever. Herein, we have expanded on this study by investigating the role of three of the plants Pithecellobium dulce (Fabaceae), Leonotis nepetifolia (Lamiaceae), and Opuntia ficus-indica (Cactaceae) on survival, fecundity, and egg viability of the dengue vector Aedes aegypti. Methods: We tested these effects using females that received a) an initial three rations of bloodmeal, and b) received no bloodmeal at all. Two controls were included; age-matched females fed on glucose solution with or without initial bloodmeal, and those fed exclusively on bloodmeal. Data was collected daily over a 30-day period. The amino acid content of Ae. aegypti guts and the amino acid content of their respective diets was detected by coupled liquid chromatography – mass spectrometry.Results: Females fed on P. dulce and exclusive bloodmeal had a shorter survival than those fed on glucose. On the other hand, females fed on L. nepetifolia survived longer than those fed exclusively on bloodmeal, whereas those fed on O. ficus-indica had the shortest survival time. With initial bloodmeal, females fed on L. nepetifolia laid 1.6-fold more eggs while those fed on the other diets laid fewer eggs, compared to those fed exclusively on bloodmeal. Hatching rates of the eggs laid varied with the diet. Mass spectroscopic analysis of gut contents of mosquitoes exposed to the different diets showed qualitative and quantitative differences in their amino acid levels. Conclusion: Our findings highlight the central role of plant nutrients in the reproductive fitness of dengue vectors which may impact their disease transmission potential.


2020 ◽  
Author(s):  
Vincent Odhiambo Nyasembe ◽  
David P. Tchouassi ◽  
Martha N. Muturi ◽  
Christian W. W. Pirk ◽  
Catherine L. Sole ◽  
...  

Abstract Background In a recent study using DNA barcoding, we identified the plants fed upon by four Afro-tropical mosquito species that vector dengue, malaria, and Rift Valley fever. Herein, we have expanded on this study by investigating the role of three of the plants Pithecellobium dulce (Fabaceae), Leonotis nepetifolia (Lamiaceae), and Opuntia ficus-indica (Cactaceae) on survival, fecundity, and egg viability of the dengue vector Aedes aegypti. Methods We tested these effects using females that received a) an initial three rations of bloodmeal, and b) received no bloodmeal at all. Two controls were included; age-matched females fed on glucose solution with or without initial bloodmeal, and those fed exclusively on bloodmeal. Data was collected daily over a 30-day period. The amino acid content of Ae. aegypti guts and the amino acid content of their respective diets was detected by coupled liquid chromatography – mass spectrometry. Results Females fed on P. dulce and exclusive bloodmeal had a shorter survival than those fed on glucose. On the other hand, females fed on L. nepetifolia survived longer than those fed exclusively on bloodmeal, whereas those fed on O. ficus-indica had the shortest survival time. With initial bloodmeal, females fed on L. nepetifolia laid 1.6-fold more eggs while those fed on the other diets laid fewer eggs, compared to those fed exclusively on bloodmeal. Hatching rates of the eggs laid varied with the diet. Mass spectroscopic analysis of gut contents of mosquitoes exposed to the different diets showed qualitative and quantitative differences in their amino acid levels. Conclusion Our findings highlight the central role of plant nutrients in the reproductive fitness of dengue vectors which may impact their disease transmission potential.


2006 ◽  
Vol 397 (1) ◽  
pp. 69-76 ◽  
Author(s):  
Li-Chiun Lee ◽  
Ya-Lin Lee ◽  
Ruey-Jyh Leu ◽  
Jei-Fu Shaw

Escherichia coli TAP (thioesterase I, EC 3.1.2.2) is a multifunctional enzyme with thioesterase, esterase, arylesterase, protease and lysophospholipase activities. Previous crystal structural analyses identified its essential amino acid residues as those that form a catalytic triad (Ser10-Asp154-His157) and those involved in forming an oxyanion hole (Ser10-Gly44-Asn73). To gain an insight into the biochemical roles of each residue, site-directed mutagenesis was employed to mutate these residues to alanine, and enzyme kinetic studies were conducted using esterase, thioesterase and amino-acid-derived substrates. Of the residues, His157 is the most important, as it plays a vital role in the catalytic triad, and may also play a role in stabilizing oxyanion conformation. Ser10 also plays a very important role, although the small residual activity of the S10A variant suggests that a water molecule may act as a poor substitute. The water molecule could possibly be endowed with the nucleophilic-attacking character by His157 hydrogen-bonding. Asp154 is not as essential compared with the other two residues in the triad. It is close to the entrance of the substrate tunnel, therefore it predominantly affects substrate accessibility. Gly44 plays a role in stabilizing the oxyanion intermediate and additionally in acyl-enzyme-intermediate transformation. N73A had the highest residual enzyme activity among all the mutants, which indicates that Asn73 is not as essential as the other mutated residues. The role of Asn73 is proposed to be involved in a loop75–80 switch-move motion, which is essential for the accommodation of substrates with longer acyl-chain lengths.


RSC Advances ◽  
2016 ◽  
Vol 6 (89) ◽  
pp. 86650-86662 ◽  
Author(s):  
Manish K. Tiwari ◽  
Phool C. Mishra

The catalytic role of iron-superoxide dismutase (Fe-SOD) in the working of ascorbic acid (AA) as a superoxide radical anion scavenger has been studied by employing a model developed recently for the active site of the enzyme.


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