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1.
Hydrogen peroxide (H2O2)-induced aggregation of calf platelets and its modification by agents with specific properties were characterized employing
a spectrophotometric assay. An Arrhenius activation energy of 20 ± 1 kcal/mol was found in the temperature range of 25‡-36‡C.
Rate inhibition occurred on either side of this temperature range, and under anaerobic conditions. Exogenous Ca2+ ions were not required but Ca2+ ions, at 1 mM-concentration, optimally increased rates and extent of aggregation at suboptimal H2O2 concentrations but only extent of aggregation at optimal H2O2 concentrations. Ba2+, Sr2+, Cd2+, Mn2+ and Ni2+ ions (1 mM) and Zn2+, Pb2+ and Hg2+ ions (10 mM) were inhibitory. The cyclo-oxygenase inhibitor, indomethacin (10-30 mM) exerted only mild inhibition by a competitive
mechanism. Another cyclo-oxygenase inhibitor, aspirin, functioned to increase aggregation. Ligands acting directly at the
prostaglandin H2/thromboxane A, receptor (5Z. 9, 11, 13E, 15(S) 15-hydroxy 9(11) epoxy methano prosta 5, 13-dien-1-oic acid, pinane thromboxane
A2, arachidonic acid, eicosapentaenoic acid, and N-ethylmaleimide) functioned as competitive inhibitors. Another platelet-activating
sulphydryl reagent, thimerosal, also inhibited competitively while the protein kinase C inhibitor, sphingosine, and the protein
kinase C modulator, Zn2+ ions, inhibited by different mechanisms. The results indicate direct action of H2O2 at the prostaglandin H2/thromboxane A2 receptor, possibly its sulphydryls, to activate the protein kinase C pathway, independently of cyclo-oxygenase products.
The results underscored the power of the kinetic approach for investigating mechanisms of platelet activation. 相似文献
2.
In cells, mitochondria, endoplasmic reticulum, and peroxisomes are the major sources of reactive oxygen species (ROS) under physiological and pathophysiological conditions. Cytochrome c (cyt c) is known to participate in mitochondrial electron transport and has antioxidant and peroxidase activities. Under oxidative or nitrative stress, the peroxidase activity of Fe3+cyt c is increased. The level of NADH is also increased under pathophysiological conditions such as ischemia and diabetes and a concurrent increase in hydrogen peroxide (H2O2) production occurs. Studies were performed to understand the related mechanisms of radical generation and NADH oxidation by Fe3+cyt c in the presence of H2O2. Electron paramagnetic resonance (EPR) spin trapping studies using 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) were performed with NADH, Fe3+cyt c, and H2O2 in the presence of methyl-β-cyclodextrin. An EPR spectrum corresponding to the superoxide radical adduct of DMPO encapsulated in methyl-β-cyclodextrin was obtained. This EPR signal was quenched by the addition of the superoxide scavenging enzyme Cu,Zn-superoxide dismutase (SOD1). The amount of superoxide radical adduct formed from the oxidation of NADH by the peroxidase activity of Fe3+cyt c increased with NADH and H2O2 concentration. From these results, we propose a mechanism in which the peroxidase activity of Fe3+cyt c oxidizes NADH to NAD•, which in turn donates an electron to O2, resulting in superoxide radical formation. A UV-visible spectroscopic study shows that Fe3+cyt c is reduced in the presence of both NADH and H2O2. Our results suggest that Fe3+cyt c could have a novel role in the deleterious effects of ischemia/reperfusion and diabetes due to increased production of superoxide radical. In addition, Fe3+cyt c may play a key role in the mitochondrial “ROS-induced ROS-release” signaling and in mitochondrial and cellular injury/death. The increased oxidation of NADH and generation of superoxide radical by this mechanism may have implications for the regulation of apoptotic cell death, endothelial dysfunction, and neurological diseases. We also propose an alternative electron transfer pathway, which may protect mitochondria and mitochondrial proteins from oxidative damage. 相似文献
3.
Xiuli Hu Wei Wang Chunqi Li Jianhua Zhang Fan Lin Aying Zhang Mingyi Jiang 《Plant Growth Regulation》2008,55(3):183-198
Here we examined whether Ca2+/Calmodulin (CaM) is involved in abscisic acid (ABA)-induced antioxidant defense and the possible relationship between CaM
and H2O2 in ABA signaling in leaves of maize (Zea mays L.) plants exposed to water stress. An ABA-deficient mutant vp5 and its wild type were used for the experimentation. We found that water stress enhanced significantly the contents of CaM
and H2O2, and the activities of chloroplastic and cytosolic superoxide dismutase (SOD), ascorbate peroxidase (APX) and glutathione
reductase (GR), and the gene expressions of the CaM1, cAPX, GR1 and SOD4 in leaves of wild-type maize. However, the increases mentioned above were almost arrested in vp5 plants and in the wild-type plants pretreated with ABA biosynthesis inhibitor tungstate (T), suggesting that ABA is required
for water stress-induced H2O2 production, the enhancement of CaM content and antioxidant defense. Besides, we showed that the up-regulation of water stress-induced
antioxidant defense was almost completely blocked by pretreatment with Ca2+ inhibitors, CaM antagonists and reactive oxygen (ROS) manipulators. Moreover, the analysis of time course of CaM and H2O2 production under water stress showed that the increase in CaM content preceded that of H2O2. These results suggested that Ca2+/CaM and H2O2 were involved in the ABA-induced antioxidant defense under water stress, and the increases of Ca2+/CaM contents triggered H2O2 production, which inversely affected the contents of CaM. Thus, a cross-talk between Ca2+/CaM and H2O2 may play a pivotal role in the ABA signaling. 相似文献
4.
Oxidation of vanadyl sulfate by H2O2 involves multiple reactions at neutral pH conditions. The primary reaction was found to be oxidation of V(IV) to V(V) using 0.5 equivalent of H2O2, based on the loss of blue color and the visible spectrum. The loss of V(IV) and formation V(V) compounds were confirmed by ESR and51V-NMR spectra, respectively. In the presence of excess H2O2 (more than two equivalents), the V(V) was converted into diperoxovanadate, the major end-product of these reactions, identified by changes in absorbance in ultraviolet region and by the specific chemical shift in NMR spectrum. The stoichiometric studies on the H2O2 consumed in this reaction support the occurrence of reactions of two-electron oxidation followed by complexing two molecules of H2O2. Addition of a variety of compounds—Tris, ethanol, mannitol, benzoate, formate (hydroxyl radical quenching), histidine, imidazole (singlet oxygen quenching), and citrate—stimulated a secondary reaction of oxygen-consumption that also used V(IV) as the reducing source. This reaction requires concomitant oxidation of vanadyl by H2O2, favoured at low H2O2:V(IV) ratio. Another secondary reaction of oxygen release was found to occur during vanadyl oxidation by H2O2 in acidic medium in which the end-product was not diperoxovanadate but appears to be a mixture of VO
3
+
(–546 ppm), VO3+ (–531 ppm) and VO
2
+
(–512 ppm), as shown by the51V-NMR spectrum. This reaction also occurred in phosphate-buffered medium but only on second addition of vanadyl. The compounds that stimulated the oxygen-consumption reaction were found to inhibit the oxygen-release reaction. A combination of these reactions occur depending on the proportion of the reactants (vanadyl and H2O2), the pH of the medium and the presence of some compounds that affect the secondary reactions. 相似文献
5.
Christian Cortés-Rojo Elizabeth Calderón-Cortés Mónica Clemente-Guerrero Salvador Manzo-Ávalos Salvador Uribe Istvan Boldogh 《Free radical research》2013,47(11):1212-1223
The deleterious effects of H2O2 on the electron transport chain of yeast mitochondria and on mitochondrial lipid peroxidation were evaluated. Exposure to H2O2 resulted in inhibition of the oxygen consumption in the uncoupled and phosphorylating states to 69% and 65%, respectively. The effect of H2O2 on the respiratory rate was associated with an inhibition of succinate-ubiquinone and succinate-DCIP oxidoreductase activities. Inhibitory effect of H2O2 on respiratory complexes was almost completely recovered by β-mercaptoethanol treatment. H2O2 treatment resulted in full resistance to QO site inhibitor myxothiazol and thus it is suggested that the quinol oxidase site (QO) of complex III is the target for H2O2. H2O2 did not modify basal levels of lipid peroxidation in yeast mitochondria. However, H2O2 addition to rat brain and liver mitochondria induced an increase in lipid peroxidation. These results are discussed in terms of the known physiological differences between mammalian and yeast mitochondria. 相似文献
6.
M. Aktar Ali Fumihiko Yasui Seiichi Matsugo Tetsuya Konishi 《Free radical research》2013,47(5):429-438
The effect of lactic acid (lactate) on Fenton based hydroxyl radical (·OH) production was studied by spin trapping, ESR, and fluorescence methods using DMPO and coumarin-3-carboxylic acid (3-CCA) as the ·OH traps respectively. The ·OH adduct formation was inhibited by lactate up to 0.4mM (lactate/iron stoichiometry = 2) in both experiments, but markedly enhanced with increasing concentrations of lactate above this critical concentration. When the H2O2 dependence was examined, the DMPO-OH signal was increased linearly with H2O2 concentration up to 1 mM and then saturated in the absence of lactate. In the presence of lactate, however, the DMPO-OH signal was increased further with higher H2O2 concentration than 1 mM, and the saturation level was also increased dependent on lactate concentration. Spectroscopic studies revealed that lactate forms a stable colored complex with Fe3+ at lactate/Fe3+ stoichiometry of 2, and the complex formation was strictly related to the DMPO-OH formation. The complex formation did not promote the H2O2 mediated Fe3+ reduction. When the Fe3+-lactate (1:2) complex was reacted with H2O2, the initial rate of hydroxylated 3-CCA formation was linearly increased with H2O2 concentrations. All the data obtained in the present experiments suggested that the Fe3+-lactate (1:2) complex formed in the Fenton reaction system reacts directly with H2O2 to produce additional ·OH in the Fenton reaction by other mechanisms than lactate or lactate/Fe3+ mediated promotion of Fe3+/Fe2+ redox cycling. 相似文献
7.
He-Qing Huang Liang-Shu Xu Feng-Zhang Zhang Xue-Hui Qiu Qing-Mei Lin Jiang-Wei Huang Hong Zao Nan-Chang Huang Run-Ying Zeng Ding Zeng 《Journal of Protein Chemistry》1998,17(1):45-52
Bacterial ferritin from Azotobacter vinelandii (AvBFo has a function in H2 uptake. The Fe3+ reduction on the surface of the iron core from AvBFo is accompanied simultaneously by H2 uptake, with a maximum activity of H2 uptake of 450 H2/AvBFo. A reduction potential of –402 mV for iron reduction on the surface of the core is found. A shift to the red the protein absorbance peaks ranging from 280 to 290 nm is observed between pH5 and 9 under 100% H2 reduction. The reduction potential for iron release becomes negative at a rate of 0.025 mV/Fe2+ released. The kinetics of iron release on the surface of the core is a first-order reaction. 相似文献
8.
Meera Rau Milind S. Patole S. Vijaya C. K. Ramakrishna Kurup T. Ramasarmal 《Molecular and cellular biochemistry》1987,75(2):151-159
Addition of vanadate, stimulated oxidation of NADH by rat liver microsomes. The products were NAD+ and H2O2. High rates of this reaction were obtained in the presence of phosphate buffer and at low pH values. The yellow-orange colored polymeric form of vanadate appears to be the active species and both ortho- and meta-vanadate gave poor activities even at mM concentrations.The activity as measured by oxygen uptake was inhibited by cyanide, EDTA, mannitol, histidine, ascorbate, noradrenaline, adriamycin, cytochrome c, Mn2+, superoxide dismutase, horseradish peroxidase and catalase. Mitochondrial outer membranes possess a similar activity of vanadate-stimulated NADH oxidation. But addition of mitochondria and some of its derivative particles abolished the microsomal activity. In the absence of oxygen, disappearance of NADH measured by decrease in absorbance at 340 nm continued at nearly the same rate since vanadate served as an electron acceptor in the microsomal system. Addition of excess catalase or SOD abolished the oxygen uptake while retaining significant rates of NADH disappearance indicating that the two activities are delinked. A mechanism is proposed wherein oxygen receives the first electron from NAD radical generated by oxidation of NADH by phosphovanadate and the consequent reduced species of vanadate (Viv) gives the second electron to superoxide to reduce it H2O2. This is applicable to all membranes whereas microsomes have the additional capability of reducing vanadate. 相似文献
9.
Among various metal ions of physiological interest, Cu2+ is uniquely capable of catalyzing the oxidation of NADH by H2O2. This oxidation is stimulated about fivefold in the presence of imidazole. A similar activating effect is found for some
imidazole derivatives (1-methyl imidazole, 2-methyl imidazole, andN-acetyl-L-histidine). Some other imidazole-containing compounds (L-histidine,L-histidine methyl ester, andL-carnosine), however, inhibit the Cu2+-catalyzed peroxidation of NADH. Other chelating agents such as EDTA andL-alanine are also inhibitory. Stoichiometry for NADH oxidation per mole of H2O2 utilized is 1, which excludes the possibility of a two-step oxidation mechanism with a nucleotide free-radical intermediate.
About 92% of the NADH oxidation product can be identified as enzymatically active NAD+. D2O, 2,5-dimethylfuran, and 1,4-diazabicyclo [2.2.2]-octane have no significant effect on the oxidation, thus excluding1O2 as a mediator. Similarly, OH· is also not a likely intermediate, since the system is not affected by various scavengers of
this radical. The results suggest that a copper-hydrogen peroxide intermediate, when complexed with suitable ligands, can
generate still another oxygen species much more reactive than its parent compound, H2O2. 相似文献
10.
Barja G 《Journal of bioenergetics and biomembranes》2002,34(3):227-233
Adequate methods to measure the rate of mitochondrial oxygen radical generation are needed since oxygen radicals are involved in many pathologies. A fluorometric method appropriate to measure the rate of generation of H2O2 in intact mitochondria is described. Just after isolation of functional mitochondria from fresh tissues, rates of generation of H2O2 are kinetically measured by fluorometry in the presence of homovanillic acid and horseradish peroxidase. The method is specific for H2O2 and is sensitive enough to assay mitochondrial H2O2 generation in the presence of respiratory substrate without inhibitors of the respiratory chain. Simultaneous measurement of mitochondrial oxygen consumption allows calculation of the free radical leak: the percentage of electrons out of sequence which reduce oxygen to oxygen radicals along the mitochondrial respiratory chain instead of reducing oxygen to water at the terminal cytochrome oxidase. The method shows instantaneous response to H2O2. This makes it appropriate to study the quick effects of different inhibitors and modulators on the rate of mitochondrial oxygen radical production. Its application to the localization of the sites where caloric restriction decreases mitochondrial oxygen radical generation in heart mitochondria is described. 相似文献
11.
Dai-Eun Sok 《Journal of neurochemistry》1999,72(1):355-362
Abstract : Alkaline phosphatase, one of the enzymes responsible for the conversion of phosphocholine into choline, was purified from bovine brain membrane, where the phosphatase is bound as glycosylphosphatidylinositollinked protein, and subjected to oxidative inactivation. The phosphatase activity, based on the hydrolysis of p-nitrophenyl phosphate and phosphocholine, decreased slightly after the exposure to H2O2. Inclusion of Cu2+ in the incubation with 1 mM H2O2 led to a rapid decrease of activity in a time- and concentration-dependent manner. In comparison, the H2O2/Cu2+ system was much more effective than the H2O2/Fe2+ system in inactivating brain phosphatase. In a further study, it was observed that the hydroxy radical scavengers mannitol, ethanol, or benzoate failed to prevent against H2O2/Cu2+-induced inactivation of the phosphatase, excluding the involvement of extraneous hydroxy radicals in metalcatalyzed oxidation. In addition, it was found that both substrates, p-nitrophenyl phosphate and phosphocholine, and an inhibitor, phosphate ion, at their saturating concentrations exhibited a remarkable, although incomplete, protection against the inactivating action of H2O2/Cu2+. A similar protection was also expressed by divalent metal ions such as Mg2+ or Mn2+. Separately, it was found that H2O2/Fe2+-induced inactivation was prevented by p-nitrophenyl phosphate or Mg2+ but not phosphate ions. Thus, it is implied that phosphocholine-hydrolyzing alkaline phosphatase in brain membrane might be one of enzymes susceptible to metal-catalyzed oxidation. 相似文献
12.
Multifunctional poly(glycidyl methacrylate) (PGMA) microspheres containing magnetic, fluorescent, and cancer cell-specific moieties were prepared in four steps: (i) preparation of parent PGMA microspheres by dispersion polymerization and their reaction with ethylenediamine to obtain amino groups, (ii) precipitation of iron ions (Fe2+ and Fe3+) to form Fe3O4 nanoparticles within the microspheres, (iii) consecutive reactions of folic acid with the amino groups on PGMA, and (iv) incorporation of fluorescein isothiocyanate into the microspheres. The microspheres were superparamagnetic, highly monodispersive, intensively fluorescent, and capable of recognizing and binding cancer cells that overexpress folic acid receptors. It was demonstrated that with these microspheres, HeLa cells could be captured from their suspension and easily moved in the direction of the externally applied magnetic field. 相似文献
13.
Oxygen consumption by alternative oxidase (AOX), present in mitochondria of many angiosperms, is known to be cyanide-resistant in contrast to cytochrome oxidase. Its activity in potato tuber (Solanum tuberosum L.) was induced following chilling treatment at 4 °C. About half of the total O2 consumption of succinate oxidation in such mitochondria was found to be sensitive to SHAM, a known inhibitor of AOX activity. Addition of catalase to the reaction mixture of AOX during the reaction decreased the rate of SHAM-sensitive oxygen consumption by nearly half, and addition at the end of the reaction released nearly half of the consumed oxygen by AOX, both typical of catalase action on H2O2. These findings with catalase suggest that the product of reduction of AOX is H2O2 and not H2O, as previously surmised. In potatoes subjected to chill stress (4 °C) for periods of 3, 5 and ?8 days the activity of AOX in mitochondria increased progressively with a corresponding increase in the AOX protein detected by immunoblot of the protein. 相似文献
14.
Anna Kärkönen Tino Warinowski Teemu H. Teeri Liisa Kaarina Simola Stephen C. Fry 《Planta》2009,230(3):553-567
A cell culture of Picea abies (L.) Karst. was used for studies of H2O2 generation during constitutive extracellular lignin formation and after elicitation by cell wall fragments of a pathogenic
fungus, Heterobasidium parviporum. Stable, micromolar levels of H2O2 were present in the culture medium during lignin formation. Elicitation induced a burst of H2O2, peaking at ca. 90 min after elicitation. Of exogenous reducing substrates that may be responsible for the synthesis of H2O2 from O2, NADH stimulated H2O2 production irrespective of elicitation. Cysteine (Cys) and glutathione (GSH) partially scavenged the constitutive H2O2, but usually increased or prolonged elicitor-induced H2O2 formation. Culture medium peroxidases were not able to generate H2O2 in vitro with Cys or GSH as reductants. These thiols, however, generated H2O2 non-enzymically at pH 4.5. [35S]Sulphate feeding to spruce cells showed that endogenous sulphur-containing compounds (including GSH, GSSG and cysteic acid)
existed in the culture medium. The apoplastic levels of these were, however, undetectable by the monobromobimane method suggesting
that their contribution to apoplastic H2O2 formation is probably minor. Azide, an inhibitor of haem-containing enzymes, slightly inhibited constitutive H2O2 generation but strongly delayed the elicitor-induced H2O2 accumulation. Diphenylene iodonium, an inhibitor of flavin-containing enzymes, efficiently inhibited H2O2 production irrespective of elicitation. Elicitation led to downregulation of the expression of several peroxidase genes,
and peroxidase activity in the culture medium was slightly reduced. Expression of three other peroxidase genes and a respiratory burst oxidase homologue (rboh) gene were upregulated. These data suggest that both peroxidases and rboh may contribute to H2O2 generation.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献
15.
O. Cantoni G. Brandi G.F. Schiavano A. Albano F. Cattabeni 《Chemico-biological interactions》1989,70(3-4)
The toxicity of H2O2 in Escherichia coli wild type and superoxide dismutase mutants was investigated under different experimental conditions. Cells were either grown aerobically, and then treated in M9 salts or K medium, or grown anoxically, and then treated in K medium. Results have demonstrated that the wild type and superoxide dismutase mutants display a markedly different sensitivity to both modes of lethality produced by H2O2 (i.e. mode one killing, which is produced by concentrations of H2O2 lower than 5 mM, and mode two killing which results from the insult generated by concentrations of H2O2 higher than 10 mM). Although the data obtained do not clarify the molecular basis of H2O2 toxicity and/or do not explain the specific function of superoxide ions in H2O2-induced bacterial inactivation, they certainly demonstrate that the latter species plays a key role in both modes of H2O2 lethality. A mechanism of H2O2 toxicity in E. coli is proposed, involving the action of a hypothetical enzyme which should work as an O2-• generating system. This enzyme should be active at low concentrations of H2O2 (<5 mM) and high concentrations of the oxidant (>5 mM) should inactivate the same enzyme. Superoxide ions would then be produced and result in mode one lethality. The resistance at intermediate H2O2 concentrations may be dependent on the inactivation of such enzyme with no superoxide ions being produced at levels of H2O2 in the range 5–10 mM. Mode two killing could be produced by the hydroxyl radical in concert with superoxide ions, chemically produced via the reaction of high concentrations of H2O2 (>10 mM) with hydroxyl radicals. The rate of hydroxyl radical production may be increased by the higher availability of Fe2+ since superoxide ions may also reduce trivalent iron to the divalent form. 相似文献
16.
Yeast cytochrome c peroxidase (CCP) efficiently catalyzes the reduction of H2O2 to H2O by ferrocytochrome c in vitro. The physiological function of CCP, a heme peroxidase that is targeted to the mitochondrial intermembrane space of Saccharomyces cerevisiae, is not known. CCP1-null-mutant cells in the W303-1B genetic background (ccp1Δ) grew as well as wild-type cells with glucose, ethanol, glycerol or lactate as carbon sources but with a shorter initial doubling time. Monitoring growth over 10 days demonstrated that CCP1 does not enhance mitochondrial function in unstressed cells. No role for CCP1 was apparent in cells exposed to heat stress under aerobic or anaerobic conditions. However, the detoxification function of CCP protected respiring mitochondria when cells were challenged with H2O2. Transformation of ccp1Δ with ccp1W191F, which encodes the CCPW191F mutant enzyme lacking CCP activity, significantly increased the sensitivity to H2O2 of exponential-phase fermenting cells. In contrast, stationary-phase (7-day) ccp1Δ-ccp1W191F exhibited wild-type tolerance to H2O2, which exceeded that of ccp1Δ. Challenge with H2O2 caused increased CCP, superoxide dismutase and catalase antioxidant enzyme activities (but not glutathione reductase activity) in exponentially growing cells and decreased antioxidant activities in stationary-phase cells. Although unstressed stationary-phase ccp1Δ exhibited the highest catalase and glutathione reductase activities, a greater loss of these antioxidant activities was observed on H2O2 exposure in ccp1Δ than in ccp1Δ-ccp1W191F and wild-type cells. The phenotypic differences reported here between the ccp1Δ and ccp1Δ-ccp1W191F strains lacking CCP activity provide strong evidence that CCP has separate antioxidant and signaling functions in yeast. 相似文献
17.
The nonenzymatic reaction of glyoxylate and H2O2 was measured under physiological conditions of the pH and concentrations of reactants. The reaction of glyoxylate and H2O2 was secondorder, with a rate constant of 2.27 l mol-1 s-1 at pH 8.0 and 25° C. The rate constant increased by 4.4 times in the presence of Zn2+ and doubled at 35°C. We propose a mechanism for the reaction between glyoxylate and H2O2. From a comparison of the rates of H2O2 decomposition by catalase and the reaction with glyoxylate, we conclude that H2O2 produced during glycolate oxidation in peroxisomes is decomposed by catalase but not by the reaction with glyoxylate, and that photorespiratory CO2 originates from glycine, but not from glyoxylate, in C3 plants. Simulation using the above rate constant and reported kinetic parameters leads to the same conclusion, and also makes it clear that alanine is a satisfactory amino donor in the conversion of glyoxylate to glycine. Some serine might be decomposed to give glycine and methylene-tetrahydrofolate; the latter is ultimately oxidized to CO2. In the simulation of the glycolate pathway of Euglena, the rate constant was high enough to ensure the decarboxylation of glyoxylate by H2O2 to produce photorespiratory CO2 during the glycolate metabolism of this organism.Abbreviations Chl
chlorophyll
- GGT
glutamate: glyoxylate aminotransferase (EC 2.6.1.4)
- Hepes
4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid
- SGT
serine: glyoxylate aminotransferase (EC 2.6.1.45)
This is the ninth in a series on the metabolism of glycolate in Euglena gracilis. The eighth is Yokota et al. (1982) 相似文献
18.
The ATP.Mg-dependent type 1 protein phosphatase is inactive as isolated but can be activated in several different ways. In this report, we show that the phosphatase can also be activated by the Fe2+/ascorbate system. Activation of the phosphatase requires both Fe2+ ion and ascorbate and the level of activation is dependent on the concentrations of Fe2+ ion and ascorbate. In the presence of 20 mM ascorbate, the Fe2+ ion concentrations required for half-maximal and maximal activation are about 0.3 and 3mM, respectively. Several common divalent metal ions, including Co2+, Ni2+, Cu2+, Mg2+, and Ca2+ ions, cannot cooperate with ascorbate to activate the phosphatase, and SH-containing reducing agents such as 2-mercaptoethanol and dithiothreitol cannot cooperate with Fe2+ ion to activate the phosphatase, indicating that activation of the phosphatase by the Fe2+/ascorbate system is a specific process. Moreover, H2O2, a strong oxidizer, could significantly diminish the phosphatase activation by the Fe2+/ascorbate system, suggesting that reduction mechanism other than SH-SS interchange is a prerequisite for the Fe2+/ascorbate-mediated phosphatase activation. Taken together, the present study provides initial evidence for a new mode of type 1 protein phosphatase activation mechanism.Abbreviations MAPK
mitogen-activated protein kinase
- MCO
metal ion-catalyzed oxidation
- kinase FA
the activating factor of ATP.Mg-dependent protein phosphatase
- I2
inhibitor-2
- EDTA
ethylenediaminetetraacetic acid
- MBP
myelin basic protein 相似文献
19.
《Bioscience, biotechnology, and biochemistry》2013,77(12):2525-2530
Chlorogenic acid is the major polyphenol in foods derived from plants and is a good substrate for polyphenol oxidase. Chlorogenic acid quinone (CQA-Q), which is an oxidative product of chlorogenic acid by polyphenol oxidase, is an important intermediate compound in enzymatic browning. CQA-Q was prepared, and its properties and the relationship with browning were examined. The quinone solution was yellow or orange, and its molecular absorption coefficient was estimated to be 1.7×103 for 325 nm and 9.7×102 for 400 nm in an acidic aqueous solution. Chlorogenic acid and H2O2 were spontaneously generated in the CQA-Q solution as the yellowish color of the solution gradually faded. A pale colored polymer was the major product in the reaction solution. Amino acids such as lysine and arginine added to CQA-Q solution did not repress the fading of the yellowish color of the solution. We concluded from these results that CQA-Q itself and a mixture of CQA-Q and amino acids did not form intensive brown pigments in the acidic aqueous solution. H2O2 spontaneously formed in the CQA-Q solution, and other polyphenols might have played an important role in the formation of the brown color by enzymatic browning. 相似文献
20.
Maximiliano J. Amenabar Daniel R. Colman Saroj Poudel Eric E. Roden Eric S. Boyd 《Environmental microbiology》2018,20(7):2523-2537
The thermoacidophilic Acidianus strain DS80 displays versatility in its energy metabolism and can grow autotrophically and heterotrophically with elemental sulfur (S°), ferric iron (Fe3+) or oxygen (O2) as electron acceptors. Here, we show that autotrophic and heterotrophic growth with S° as the electron acceptor is obligately dependent on hydrogen (H2) as electron donor; organic substrates such as acetate can only serve as a carbon source. In contrast, organic substrates such as acetate can serve as electron donor and carbon source for Fe3+ or O2 grown cells. During growth on S° or Fe3+ with H2 as an electron donor, the amount of CO2 assimilated into biomass decreased when cultures were provided with acetate. The addition of CO2 to cultures decreased the amount of acetate mineralized and assimilated and increased cell production in H2/Fe3+ grown cells but had no effect on H2/S° grown cells. In acetate/Fe3+ grown cells, the presence of H2 decreased the amount of acetate mineralized as CO2 in cultures compared to those without H2. These results indicate that electron acceptor availability constrains the variety of carbon sources used by this strain. Addition of H2 to cultures overcomes this limitation and alters heterotrophic metabolism. 相似文献