83 results on '"Tankred Schewe"'
Search Results
2. How do dietary flavanols improve vascular function? A position paper
- Author
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Helmut Sies, Tankred Schewe, and Yvonne Steffen
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Umbilical Veins ,Antioxidant ,Flavonols ,Endothelium ,medicine.medical_treatment ,Biophysics ,Pharmacology ,Nitric Oxide ,medicine.disease_cause ,Biochemistry ,Catechin ,Proinflammatory cytokine ,Inhibitory Concentration 50 ,Structure-Activity Relationship ,chemistry.chemical_compound ,Phenols ,medicine ,Humans ,Molecular Biology ,Cells, Cultured ,Flavonoids ,NADPH oxidase ,biology ,Superoxide ,Endothelial Cells ,NADPH Oxidases ,Polyphenols ,Diet ,Oxidative Stress ,medicine.anatomical_structure ,chemistry ,Apocynin ,biology.protein ,Endothelium, Vascular ,Peroxynitrite ,Oxidative stress - Abstract
Epidemiological and clinical studies revealed that high-flavanol diet or isolated (-)-epicatechin improves the function of the vascular endothelium, as assessed by flow-mediated dilation, through elevation of bioavailability and bioactivity of NO*. We have demonstrated that exposure of human endothelial cells to (-)-epicatechin elevates the cellular levels of NO* and cyclic GMP and protects against oxidative stress elicited by proinflammatory agonists. (-)-Epicatechin acts like a prodrug, since these effects involve O-methylation of the flavanol and are attributed to apocynin-like inhibition of endothelial NADPH oxidase. Thus, generation of superoxide and peroxynitrite is diminished and, consequently, the cellular NO* level is preserved or augmented. We propose therefore that endothelial NO* metabolism rather than general antioxidant activity is a major target of dietary flavanols and that NADPH oxidase activity is a crucial site of action. Moreover, flavonoid glucuronides appear to serve as plasma transport metabolites to target cells rather than solely as excretion products. Implications for the interpretation of the role of dietary polyphenols for cardiovascular health are discussed.
- Published
- 2008
3. Kinetic evidence for rapid oxidation of (–)-epicatechin by human myeloperoxidase
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Christa Jakopitsch, Jürgen Arnhold, Helmut Sies, Christian Obinger, Tankred Schewe, Paul G. Furtmüller, and Holger Spalteholz
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chemistry.chemical_classification ,NADPH oxidase ,biology ,Stereochemistry ,Biophysics ,Electrons ,Cell Biology ,Biochemistry ,Catechin ,Nitric oxide ,Chemical kinetics ,Kinetics ,chemistry.chemical_compound ,Enzyme ,chemistry ,Myeloperoxidase ,Apocynin ,biology.protein ,Humans ,Organic chemistry ,Oxidation-Reduction ,Molecular Biology ,Heme ,Peroxidase - Abstract
Apocynin has been reported to require dimerization by myeloperoxidase (MPO) to inhibit leukocyte NADPH oxidase. (-)-Epicatechin, a dietary flavan-3-ol, has been identified as a 'prodrug' of apocynin-like metabolites that inhibit endothelial NADPH oxidase activity and elevate the cellular level of nitric oxide. Since (-)-epicatechin has tentatively been identified as substrate of MPO, we studied the one-electron oxidation of (-)-epicatechin by MPO. By using multi-mixing stopped-flow technique, we demonstrate that (-)-epicatechin is one of the most efficient electron donors for heme peroxidases investigated so far. Second order rate constants for the (-)-epicatechin-mediated conversion of MPO-compound I to compound II and compound II to resting enzyme were estimated to be 1.9 x 10{sup 7} and 4.5 x 10{sup 6} M{sup -1} s{sup -1}, respectively (pH 7, 25 deg. C). The data indicate that (-)-epicatechin is capable of undergoing fast MPO-mediated one-electron oxidation.
- Published
- 2008
4. (–)-Epicatechin elevates nitric oxide in endothelial cells via inhibition of NADPH oxidase
- Author
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Tankred Schewe, Helmut Sies, and Yvonne Steffen
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Biophysics ,Pharmacology ,Nitric Oxide ,Methylation ,Biochemistry ,Catechin ,Umbilical vein ,Umbilical Cord ,Nitric oxide ,chemistry.chemical_compound ,Humans ,Enzyme Inhibitors ,Cyclic GMP ,Molecular Biology ,Cells, Cultured ,NADPH oxidase ,Molecular Structure ,biology ,Angiotensin II ,Acetophenones ,Endothelial Cells ,NADPH Oxidases ,Cell Biology ,Prodrug ,chemistry ,Apocynin ,cardiovascular system ,biology.protein ,Signal transduction ,Signal Transduction - Abstract
Dietary (-)-epicatechin is known to improve bioactivity of (*)NO in arterial endothelium of humans, but the mode of action is unclear. We used the fluorophore 4,5-diaminofluorescein diacetate to visualize the (*)NO level in living human umbilical vein endothelial cells (HUVEC). Untreated cells showed only a weak signal, whereas pretreatment with (-)-epicatechin (10 microM) or apocynin (100 microM) elevated the (*)NO level. The effects were more pronounced when the cells were treated with angiotensin II with or without preloading of the cells with (*)NO via PAPA-NONOate. While (-)-epicatechin scavenged O2(*-), its O-methylated metabolites prevented O2(*-) generation through inhibition of endothelial NADPH oxidase activity, even more strongly than apocynin. From the effect of 3,5-dinitrocatechol, an inhibitor of catechol-O-methyltransferase (COMT), on HUVEC it is concluded that (-)-epicatechin serves as 'prodrug' for conversion to apocynin-like NADPH oxidase inhibitors. These data indicate an (*)NO-preserving effect of (-)-epicatechin via suppression of O2(*-)-mediated loss of (*)NO.
- Published
- 2007
5. Protein modification elicited by oxidized low-density lipoprotein (LDL) in endothelial cells: Protection by (–)-epicatechin
- Author
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Yvonne Steffen, Tobias Jung, Helmut Sies, Tilman Grune, Lars-Oliver Klotz, and Tankred Schewe
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Proteasome Endopeptidase Complex ,Endothelium ,Nitric Oxide ,Biochemistry ,Catechin ,4-Hydroxynonenal ,chemistry.chemical_compound ,Enos ,Dichlorofluorescein ,Physiology (medical) ,medicine ,Animals ,Humans ,Endothelial dysfunction ,NADPH oxidase ,biology ,Chemistry ,NADPH Oxidases ,Proteins ,medicine.disease ,biology.organism_classification ,Immunohistochemistry ,Molecular biology ,Lipoproteins, LDL ,medicine.anatomical_structure ,Apocynin ,biology.protein ,Cattle ,lipids (amino acids, peptides, and proteins) ,Endothelium, Vascular ,Lipoprotein - Abstract
The action of oxidatively modified low-density lipoprotein (oxLDL) on vascular endothelial cells has been proposed to be a crucial process leading to endothelial dysfunction and atherogenesis. OxLDL was shown here to elicit oxidative stress in bovine aortic endothelial cells or human umbilical vein endothelial cells, as judged by an increase in 2',7'-dichlorofluorescein fluorescence and elevated levels of carbonylated, nitrated, and 2-hydroxynonenal-coupled proteins. These effects were sensitive to apocynin, indicating involvement of NADPH oxidase. A 170-kDa polypeptide carbonylated upon exposure of cells to oxLDL was identified by immunoprecipitation as EGF receptor. Immunocytochemical visualization by confocal microscopy revealed the highest levels of modified proteins in the perinuclear region. Exposure of endothelial cells to oxLDL led to modulation of the expression levels of *NO synthases; the endothelial isoform (eNOS) was down-regulated via proteasomal degradation, whereas the inducible isoform (iNOS) was up-regulated in an enzymatically active state. eNOS protein was found to be both carbonylated and nitrated upon exposure of cells to oxLDL. iNOS contributed to the generation of modified proteins as judged by the effects of the selective inhibitor L-NIO. These oxLDL-elicited changes in vascular endothelial cells described were suppressed by (-)-epicatechin, a dietary polyphenol, which inhibited NADPH oxidase activity in these cells.
- Published
- 2007
6. Myeloperoxidase-mediated LDL oxidation and endothelial cell toxicity of oxidized LDL: attenuation by (−)-epicatechin
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Tankred Schewe, Yvonne Steffen, and Helmut Sies
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Pharmacology ,Biochemistry ,Antioxidants ,Catechin ,Nitric oxide ,chemistry.chemical_compound ,In vivo ,medicine ,Humans ,Peroxidase ,Cacao ,Aspirin ,biology ,Endothelial Cells ,General Medicine ,Bioavailability ,Vasoprotective ,Lipoproteins, LDL ,Endothelial stem cell ,chemistry ,Myeloperoxidase ,Toxicity ,biology.protein ,lipids (amino acids, peptides, and proteins) ,Oxidation-Reduction ,medicine.drug - Abstract
Recent data suggest an inverse epidemiological association between intake of flavanol-rich cocoa products and cardiac mortality. Potential beneficial effect of cocoa may be attributed to flavanol-mediated improvement of endothelial function, as well as to enhancement of bioavailability and bioactivity of nitric oxide in vivo. ( - )-Epicatechin is one bioactive flavanol found in cocoa. This review deals with protective actions of ( - )-epicatechin on two key processes in atherogenesis, oxidation of LDL and damage to endothelial cell by oxidized LDL (oxLDL), with emphasis on data from this laboratory. ( - )-Epicatechin not only abrogates or attenuates LDL oxidation but also counteracts deleterious actions of oxLDL on vascular endothelial cells. These protective actions are only partially shared by other vasoprotective agents such as vitamins C and E or aspirin. Thus, ( - )-epicatechin appears to be a pleiotropic protectant for both LDL and endothelial cells.
- Published
- 2006
7. Epicatechin protects endothelial cells against oxidized LDL and maintains NO synthase
- Author
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Yvonne Steffen, Helmut Sies, and Tankred Schewe
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Nitric Oxide Synthase Type III ,Lactacystin ,Biophysics ,Pharmacology ,Biochemistry ,Antioxidants ,Catechin ,Cell Line ,Nitric oxide ,Pathogenesis ,chemistry.chemical_compound ,medicine ,Humans ,Cytotoxic T cell ,Endothelial dysfunction ,Molecular Biology ,DNA Primers ,Peroxidase ,Base Sequence ,biology ,Reverse Transcriptase Polymerase Chain Reaction ,Cell Biology ,medicine.disease ,Lipoproteins, LDL ,chemistry ,Proteasome ,Myeloperoxidase ,Proteasome inhibitor ,biology.protein ,Nitric Oxide Synthase ,medicine.drug - Abstract
Intake of flavanol-rich food or beverages was previously shown to ameliorate endothelial function and to enhance bioactivity of nitric oxide with individuals at risk for cardiovascular disease. Here, we examined whether the major dietary flavanol, (−)-epicatechin, counteracts the action of oxidized LDL on endothelial cells, an action considered pivotal for endothelial dysfunction in the pathogenesis of atherosclerosis. Oxidation by myeloperoxidase plus nitrite rendered human LDL cytotoxic towards endothelial cells, more so than oxidation by Cu2+. Oxidized LDL also caused a marked loss of endothelial NO synthase protein which did not occur in the presence of a proteasome inhibitor, lactacystin. Both actions of oxidized LDL, which were not evoked by native LDL, were effectively counteracted by (−)-epicatechin. We conclude that dietary flavanols contribute to protection of the integrity of endothelial cells not only by scavenging free radicals but also by maintaining endothelial NO synthase.
- Published
- 2005
8. Self-Inactivation by 13-Hydroperoxylinoleic Acid and Lipohydroperoxidase Activity of the Reticulocyte Lipoxygenase
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Tankred Schewe, Brigitte Härtel, Peter Ludwig, and Samuel M. Rapoport
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Reticulocytes ,Chemical Phenomena ,Linoleic acid ,chemistry.chemical_element ,Biochemistry ,Oxygen ,Ferrous ,Linoleic Acid ,chemistry.chemical_compound ,Sodium borohydride ,Lipoxygenase ,Reticulocyte ,medicine ,Animals ,Lipoxygenase Inhibitors ,chemistry.chemical_classification ,biology ,Chemistry ,medicine.anatomical_structure ,Enzyme ,Linoleic Acids ,chemistry ,biology.protein ,Ferric ,Rabbits ,Soybeans ,medicine.drug - Abstract
1. The self-inactivation of lipoxygenase from rabbit reticulocytes with linoleic acid at 37 degrees C is caused by the product 13-hydroperoxylinoleic acid. This inactivation is promoted by either oxygen or linoleic acid. 2. Lipohydroperoxidase activity was demonstrated with 13-hydroperoxylinoleic acid plus linoleic acid as hydrogen donor under anaerobic conditions at 2 degrees C. The products were 13-hydroxylinoleic acid, oxodienes and compounds of non-diene structure similar to those produced by soybean lipoxygenase-1. 3. 13-Hydroperoxylinoleic acid also changed the absorbance and fluorescence properties of reticulocyte lipoxygenase. The results indicate that one equivalent of 13-hydroperoxylinoleic acid converts the enzyme from the ferrous state into the ferric state as described for soybean lipoxygenase-1. The spectral changes were reversed by sodium borohydride at 2 degrees C, but not at 37 degrees C; it is assumed that the ferric form of reticulocyte lipoxygenase suffers inactivation.
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- 2005
9. Cocoa polyphenols and inflammatory mediators
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Tankred Schewe, Helmut Sies, Christian Heiss, and Malte Kelm
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Leukotrienes ,Antioxidant ,medicine.medical_treatment ,Metabolite ,Lipoxygenase ,Medicine (miscellaneous) ,Pharmacology ,Nitric Oxide ,Antioxidants ,Nitric oxide ,Proinflammatory cytokine ,Lipid peroxidation ,chemistry.chemical_compound ,medicine ,Animals ,Humans ,Enzyme Inhibitors ,Peroxidase ,Flavonoids ,Cacao ,Leukotriene ,Nutrition and Dietetics ,biology ,food and beverages ,Biochemistry ,chemistry ,Myeloperoxidase ,biology.protein ,Lipid Peroxidation ,Peroxynitrite - Abstract
Cocoa products are sources of flavan-3-ols, which have attracted interest regarding cardiovascular health. This review provides a survey of our research on the effects of cocoa polyphenols on leukotriene and nitric oxide (NO) metabolism and on myeloperoxidase-induced modification of LDL. Because intake of flavonoid-rich chocolate by human subjects was reported to decrease the plasma concentrations of proinflammatory cysteinyl leukotrienes, we assessed whether cocoa polyphenols inhibited human 5-lipoxygenase, the key enzyme of leukotriene synthesis. (-)-Epicatechin and other cocoa flavan-3-ols proved to be inhibitory at the enzyme level. This action may confer antileukotriene action in vivo. In a double-blind crossover study, 20 individuals at risk for cardiovascular diseases received cocoa beverages with high or low contents of flavan-3-ols. NO-dependent, flow-mediated dilation of the brachial artery and concentrations of nitroso compounds in plasma were measured, and it was shown that ingestion of the high-flavanol coca drink but not the low-flavanol cocoa drink significantly increased plasma concentrations of nitroso compounds and flow-mediated dilation of the brachial artery. Therefore, ingested flavonoids may reverse endothelial dysfunction through enhancement of NO bioactivity. Oxidative modification of LDL appears to be crucial for atherogenesis, and one of the mediators is the proinflammatory proatherogenic enzyme myeloperoxidase. Micromolar concentrations of (-)-epicatechin or other flavonoids were found to suppress lipid peroxidation in LDL induced by myeloperoxidase in the presence of physiologically relevant concentrations of nitrite, an NO metabolite. Adverse effects of NO metabolites, such as nitrite and peroxynitrite, were thus attenuated.
- Published
- 2005
10. Myeloperoxidase-induced lipid peroxidation of LDL in the presence of nitrite. Protection by cocoa flavanols
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Helmut Sies and Tankred Schewe
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Flavonols ,Clinical Biochemistry ,Biochemistry ,Nitric oxide ,Lipid peroxidation ,chemistry.chemical_compound ,medicine ,Humans ,Endothelial dysfunction ,Nitrite ,Nitrites ,Peroxidase ,Cacao ,biology ,Chemistry ,General Medicine ,Metabolism ,Atherosclerosis ,medicine.disease ,Lipoproteins, LDL ,Myeloperoxidase ,biology.protein ,Molecular Medicine ,Lipid Peroxidation ,Peroxynitrite ,Lipoprotein - Abstract
Lipid peroxidation (LPO) of low-density lipoprotein (LDL) is believed to be a pivotal process rendering this plasma lipoprotein atherogenic. Several endogenous factors have been proposed to mediate LPO of LDL, among them myeloperoxidase (MPO), which is active in atherosclerotic lesions, and the plasma level of which has been proposed to be a prognostic parameter for cardiac events. Nitrite, a major oxidation product of nitric oxide, is substrate of MPO and a cofactor of MPO-mediated LPO under physiological conditions. Dietary flavonoids including (-)-epicatechin, a major flavan-3-ol in cocoa products, grapes and wine, are substrates of MPO as well as potent inhibitors of LPO in LDL at micromolar concentrations. Moreover, they strongly suppress protein tyrosine nitration of LDL by MPO/nitrite or peroxynitrite. By blunting undesirable MPO-mediated actions of nitrite, presumably via scavenging of the strong prooxidant and nitrating *NO2 radical, dietary flavonoids modulate NO metabolism in a favorable direction and thus counteract endothelial dysfunction. This article gives a survey on recent progress in this field with special reference to own recently published work.
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- 2005
11. Flavanol-rich cocoa drink lowers plasma F 2 -isoprostane concentrations in humans
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Ingrid Wiswedel, Tankred Schewe, Eberhard Alexander Pfister, Siegfried Kropf, Daniela Hirsch, Martin Gruening, and Helmut Sies
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Adult ,Male ,Antioxidant ,Isoprostane ,medicine.medical_treatment ,medicine.disease_cause ,Biochemistry ,Antioxidants ,Catechin ,Beverages ,Lipid peroxidation ,chemistry.chemical_compound ,Double-Blind Method ,Physiology (medical) ,medicine ,Humans ,Ingestion ,Food science ,Exercise ,Flavonoids ,Cacao ,F2-Isoprostanes ,Cross-Over Studies ,Dose-Response Relationship, Drug ,Chemistry ,Fatty Acids ,Crossover study ,Dose–response relationship ,Postprandial ,Lipid Peroxidation ,Oxidative stress - Abstract
Flavan-3-ols are potent antioxidants in vitro, but convincing evidence for antioxidant action in vivo is lacking. We examined whether an oxidative stress-mediated increase in plasma F(2)-isoprostanes is counteracted by a flavanol-rich cocoa beverage. Twenty volunteers were examined in a comparative randomized double-blind crossover design with respect to ingestion of high-flavanol cocoa drink (HFCD; 187 mg flavan-3-ols/100 ml) vs. low-flavanol cocoa drink (LFCD; 14 mg/100 ml). With 10 individuals, the treatment was combined with strenuous physical exercise. Total (esterified plus nonesterified) F(2)-isoprostanes were analyzed by GC/MS. LFCD caused a slight increase in the mean (+/- SEM) plasma concentrations of F(2)-isoprostanes 2 and 4 h after intake (2.16 +/- 0.19 nM at 4 h vs. 1.76 +/- 0.11 nM at 0 h, n = 10), which may be attributable to postprandial oxidative stress. This increase did not occur with HFCD (1.57 +/- 0.06 nM at 4 h vs. 1.65 +/- 0.10 nM at 0 h, n = 10). The difference in F(2)-isoprostanes 2 and 4 h after intake of HFCD vs. LFCD became statistically significant when the intake was combined with physical exercise (P < 0.01, ANOVA). We conclude that dietary flavanols, using cocoa drink as example, can lower the plasma level of F(2)-isoprostanes, indicators of in vivo lipid peroxidation.
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- 2004
12. Flavonoids of Cocoa Inhibit Recombinant Human 5-Lipoxygenase
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Helmut Sies, Hartmut Kühn, and Tankred Schewe
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Leukotrienes ,Flavonoid ,Medicine (miscellaneous) ,Antioxidants ,Catechin ,Lipoxygenase ,chemistry.chemical_compound ,Escherichia coli ,Biflavonoids ,Humans ,Protein Isoforms ,Proanthocyanidins ,Lipoxygenase Inhibitors ,Flavonoids ,chemistry.chemical_classification ,Cacao ,Leukotriene ,Nutrition and Dietetics ,Dose-Response Relationship, Drug ,biology ,Hydrolysis ,Metabolism ,Leukotriene A4 ,Recombinant Proteins ,Molecular Weight ,Enzyme ,chemistry ,Biochemistry ,Proanthocyanidin ,Arachidonate 5-lipoxygenase ,biology.protein ,Leukotriene Antagonists ,Arachidonic acid - Abstract
(-)-Epicatechin and its related oligomers, the procyanidins, are present in sizable amounts in some cocoas and chocolates. Intake of flavonoid-rich chocolate in humans has been reported to increase the plasma level of (-)-epicatechin and concomitantly to significantly decrease the plasma level of proinflammatory cysteinyl leukotrienes. Because leukotrienes are formed via the 5-lipoxygenase pathway of arachidonic acid metabolism, we examined whether 5-lipoxygenase is a possible target for the flavonoids of cocoa. Recombinant human 5-lipoxygenase was reacted with arachidonic acid and yielded a mixture of mainly 5-hydroperoxy-6E,8Z, 11Z,14Z-eicosatetraenoic acid (5-HpETE) and hydrolysis products of 5,6-leukotriene A(4) (LTA(4)). The formation of these products was significantly inhibited by (-)-epicatechin in a dose-dependent manner with 50% inhibitory concentrations (IC(50)) of 22 and 50 micromol/L, respectively. Among the procyanidin fractions isolated from the seeds of Theobroma cacao, only the dimer fraction and, to a lesser extent, the trimer through pentamer fractions exhibited comparable effects, whereas the larger procyanidins (hexamer through nonamer) were almost inactive. We conclude that (-)-epicatechin and its low-molecular procyanidins inhibit both dioxygenase and LTA(4) synthase activities of human 5-lipoxygenase and that this action may contribute to a putative anti-inflammatory effect of cocoa products.
- Published
- 2002
13. Epicatechin Selectively Prevents Nitration but Not Oxidation Reactions of Peroxynitrite
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Peter Schroeder, Darius P. Buchczyk, Lars-Oliver Klotz, Helmut Sies, Tankred Schewe, and Christian D. Sadik
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Lipoxygenase ,Biophysics ,Oxidative phosphorylation ,Protein Serine-Threonine Kinases ,p38 Mitogen-Activated Protein Kinases ,Biochemistry ,Catechin ,Mice ,chemistry.chemical_compound ,Proto-Oncogene Proteins ,Nitration ,Animals ,Sulfhydryl Compounds ,Phosphorylation ,Tyrosine ,Molecular Biology ,Protein kinase B ,Cells, Cultured ,Glyceraldehyde 3-phosphate dehydrogenase ,Mitogen-Activated Protein Kinase 1 ,Mitogen-Activated Protein Kinase 3 ,Nitrates ,Cell-Free System ,Dose-Response Relationship, Drug ,biology ,Kinase ,Glyceraldehyde-3-Phosphate Dehydrogenases ,Cell Biology ,Fluoresceins ,Enzyme Activation ,chemistry ,Mitogen-activated protein kinase ,biology.protein ,Endothelium, Vascular ,Mitogen-Activated Protein Kinases ,Oxidation-Reduction ,Proto-Oncogene Proteins c-akt ,Peroxynitrite - Abstract
The flavanol (-)-epicatechin has been found to protect against damage inflicted by peroxynitrite, an inflammatory intermediate. Here, epicatechin was tested in systems of increasing complexity. The compound efficiently protected against nitration of protein tyrosine residues by peroxynitrite (IC(50) approximately 0.02 mol epicatechin/mol peroxynitrite). However, at epicatechin concentrations completely preventing nitration of tyrosine by peroxynitrite, protection against the oxidative inactivation of glyceraldehyde-3-phosphate dehydrogenase or soybean lipoxygenase-1 was marginal (IC(50)1 mol epicatechin/mol peroxynitrite), approximately two orders of magnitude less. Likewise, epicatechin was relatively ineffective against oxidation of thiols in cell lysates, and against the oxidation of 2',7'-dichlorodihydrofluorescein in cultured cells. The activation of the kinases Akt/protein kinase B, ERK1/2 and p38-MAPK by peroxynitrite in murine aorta endothelial cells was not altered by epicatechin, suggesting that activation of these kinases is due to processes other than tyrosine nitration.
- Published
- 2001
14. Phospholipase A2s and lipid peroxidation
- Author
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Tankred Schewe and Santosh Nigam
- Subjects
Reticulocytes ,Arteriosclerosis ,Phospholipid ,Biology ,Phospholipase ,Phospholipases A ,Lipid peroxidation ,Membrane Lipids ,chemistry.chemical_compound ,Animals ,Arachidonate 15-Lipoxygenase ,Humans ,Molecular Biology ,Phospholipids ,Phospholipase A ,Membranes ,Phospholipase B ,Platelet-activating factor ,Cell Membrane ,Erythrocyte Aging ,Intracellular Membranes ,Cell Biology ,1-Alkyl-2-acetylglycerophosphocholine Esterase ,Asthma ,Cell biology ,Adenosine Diphosphate ,Oxidative Stress ,chemistry ,Biochemistry ,Eicosanoids ,Calcium ,lipids (amino acids, peptides, and proteins) ,Arachidonic acid ,Lipid Peroxidation ,NADP - Abstract
Lipid peroxidation of membrane phospholipids can proceed both enzymatically via the mammalian 15-lipoxygenase-1 or the NADPH-cytochrome P-450 reductase system and non-enzymatically. In some cells, such as reticulocytes, this process is biologically programmed, whereas in the majority of biological systems lipid peroxidation is a deleterious process that has to be repaired via a deacylation-reacylation cycle of phospholipid metabolism. Several reports in the literature pinpoint a stimulation by lipid peroxidation of the activity of secretory phospholipase A(2)s (mainly pancreatic and snake venom enzymes) which was originally interpreted as a repair function. However, recent experiments from our laboratory have demonstrated that in mixtures of lipoxygenated and native phospholipids the former are not preferably cleaved by either secretory or cytosolic phospholipase A(2)s. We propose that the platelet activating factor (PAF) acetylhydrolases of type II, which cleave preferentially peroxidised or lipoxygenated phospholipids, are competent for the phospholipid repair, irrespective of their role in PAF metabolism. A corresponding role of Ca(2+)-independent phospholipase A(2), which has been proposed to be involved in phospholipid remodelling in biomembranes, has not been addressed so far. Direct and indirect 15-lipoxygenation of phospholipids in biomembranes modulates cell signalling by several ways. The stimulation of phospholipase A(2)-mediated arachidonic acid release may constitute an alternative route of the arachidonic acid cascade. Thus, 15-lipoxygenase-mediated oxygenation of membrane phospholipids and its interaction with phospholipase A(2)s may play a crucial role in the pathogenesis of diseases, such as bronchial asthma and atherosclerosis.
- Published
- 2000
15. Arachidonic acid stimulates cell growth and forms a novel oxygenated metabolite in Candida albicans11Part of the data were presented at the 6th International Conference on Eicosanoids and Other Bioactive Lipids in Cancer, Inflammation and Related Diseases, September 1999, Boston, MA, Abstract No. 67
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Johan L.F. Kock, Santosh Nigam, Tankred Schewe, Roberto Ciccoli, and Rupal Deva
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Alternative oxidase ,biology ,Cell growth ,Metabolite ,Cell Biology ,biology.organism_classification ,Corpus albicans ,Yeast ,Proinflammatory cytokine ,chemistry.chemical_compound ,chemistry ,Biochemistry ,Arachidonic acid ,Candida albicans ,Molecular Biology - Abstract
Infection of human tissues by Candida albicans has been reported to cause the release of arachidonic acid (AA), eicosanoids and other proinflammatory mediators from host cells. Therefore, we investigated the interaction of this pathogen with AA. AA stimulated cell growth at micromolar concentrations when used as a sole carbon source. Moreover, it selectively inhibited the antimycin A-resistant alternative oxidase. [1-(14)C]AA was completely metabolised by C. albicans. Only one-seventh of the radioactivity metabolised was found in CO(2), whereas two-thirds occurred in carbohydrates suggesting a predominant role of the glyoxalate shunt of citrate cycle. About 1% of radioactivity was found in polar lipids including eicosanoids. A novel AA metabolite, which revealed immunoreactivity with an antibody against 3(R)-hydroxy-oxylipins, was identified as 3, 18-dihydroxy-5,8,11,14-eicosatetraenoic acid. Using immunofluorescence microscopy, endogenous 3(R)-hydroxy-oxylipins were found in hyphae but not in yeast cells. Such compounds have recently been shown to be connected with the sexual stage of the life cycle of Dipodascopsis uninucleata. Together, we propose that infection-mediated release of AA from host cells may modulate cell growth, morphogenesis and invasiveness of C. albicans by several modes. A better understanding of its role is thus promising for novel approaches towards the treatment of human mycoses.
- Published
- 2000
16. Biological actions of the free acid of hepoxilin A3 on human neutrophils
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Mark Sutherland, Santosh Nigam, and Tankred Schewe
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Pharmacology ,Arachidonic Acid ,Neutrophils ,Leukotriene B4 ,Chemotaxis ,In Vitro Techniques ,Pertussis toxin ,Biochemistry ,Respiratory burst ,Chemotaxis, Leukocyte ,chemistry.chemical_compound ,8,11,14-Eicosatrienoic Acid ,chemistry ,Eicosanoid ,Hepoxilin ,Humans ,Liberation ,Arachidonic acid ,Signal Transduction - Abstract
In earlier reports and reviews, it was suggested that unlike its methyl ester, the free acid form of the 12-lipoxygenase-derived eicosanoid hepoxilin A 3 (HXA 3 ) does not enter neutrophils and other cells. Therefore, in the past, most studies on the biological activities of HXA 3 on human neutrophils were conducted with its methyl ester. Here, we present evidence that free HXA 3 is biologically active towards human neutrophils at submicromolar concentrations, which may occur under certain circumstances in vivo . Thus, HXA 3 caused chemotaxis at concentrations as low as 30–40 nM, an effect which was attenuated at higher concentrations of this eicosanoid. Its chemotactic potency proved to be comparable to that of leukotriene B 4 , but higher than that of the chemotactic peptide formyl-methionyl-leucyl-phenylalanine (fMLP), and greatly exceeded that of the other 12-lipoxygenase metabolite, 12( S )-hydroxy-5,8,10,14-eicosatetraenoic acid, which was inactive at comparable concentrations. The chemotactic activity of HXA 3 was not abolished by serum albumin, but it was suppressed by pertussis toxin. Unlike fMLP, at this concentration range HXA 3 did not cause respiratory burst or aggregation of the neutrophils or activation of protein kinase C. These observations suggest a remarkably selective and specific receptor-mediated process. At concentrations higher than 1 μM, HXA 3 gives rise to an instantaneous release of calcium from intracellular stores which causes, however, only a slight, if any, liberation of arachidonic acid. On the other hand, pretreatment of the neutrophils with submicromolar concentrations of HXA 3 significantly blunts the liberation of arachidonic acid caused by fMLP.
- Published
- 2000
17. [Untitled]
- Author
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T. Strauss, Santosh Nigam, D. P. Smith, Tankred Schewe, J.L.F. Kock, I. Paul, Anna-Maria Botha, and D. Linke
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Mucor ,Mucorales ,biology ,General Medicine ,Oxylipin ,biology.organism_classification ,Microbiology ,Rhizomucor ,Absidia ,Cunninghamella ,Botany ,Subgenus ,Mortierella ,Molecular Biology - Abstract
The distribution of endogenous 3-hydroxylipins (3-OH oxylipins) in representatives of the Mucorales was mapped using immunofluorescence microscopy. Strains of each of the following genera were examined: Absidia, Actinomucor, Cunninghamella, Mortierella (subgenus Micromucor), Mortierella (subgenus Mortierella), Mucor and Rhizomucor. Immunofluorescence microscopy was carried out using an antibody that was raised against 3R-hydroxy-5Z,8Z,11Z,14Z-eicosatetraenoic acid (3R-HETE), which cross-reacts with other 3-OH oxylipins. Subsequently, the occurrence and distribution of the antibody on the various reproductive stages of each fungus was noted. In Absidia, Actinomucor, Mortierella (subgenus Micromucor), Mucor and Rhizomucor, 3-OH oxylipins were found to be associated with the columellae and/or wall of the sporangium. In the representative of Cunninghamella, the 3-OH oxylipins were associated with the single-spored sporangiola. No 3-OH oxylipins were detected in the strains representing Mortierella (subgenus Mortierella).
- Published
- 2000
18. Metabolic suppression of platelet-type 12-lipoxygenase in human uterine cervix with invasive carcinoma
- Author
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Yoshikazu Yamasaki, Mark Sutherland, Santosh Nigam, Hiroshi Ikawa, Tankred Schewe, G. Sravan Kumar, Natsuo Ueda, and Shozo Yamamoto
- Subjects
Blood Platelets ,Cancer Research ,medicine.medical_specialty ,Cervix Uteri ,Biology ,Arachidonate 12-Lipoxygenase ,medicine.disease_cause ,Substrate Specificity ,Endometrium ,Western blot ,Internal medicine ,medicine ,Carcinoma ,Humans ,Neoplasm Invasiveness ,12-Hydroxy-5,8,10,14-eicosatetraenoic Acid ,Cervix ,chemistry.chemical_classification ,medicine.diagnostic_test ,Cancer ,medicine.disease ,Molecular biology ,Epithelium ,Genes, bcl-2 ,Endocrinology ,Enzyme ,medicine.anatomical_structure ,Oncology ,chemistry ,Uterine Neoplasms ,Myometrium ,Immunohistochemistry ,Female ,Carcinogenesis - Abstract
Several types of lipoxygenases with various functions occur in mammalian cells. Although the presence of 12-lipoxygenase activity has been reported in uterine tissues, neither its type nor its biological functions have yet been established. Moreover, the putative role of uterine 12-lipoxygenase in cervical cancer has not been addressed before. Homogenates of uterine tissues from women without cancer and from patients with invasive cervical carcinoma were incubated with (1-14C)-arachidonic acid under various conditions and the labelled reaction products were analyzed both by thin-layer chromatography and by high-pressure liquid chromatography. 12-Lipoxygenase protein was estimated by Western blot using anti-serum against recombinant human platelet-type 12-lipoxygenase. Highest concentrations and activities of 12-lipoxygenase were found in the exocervix. The formation of 12S-hydroxy-5Z,8Z,10E,14Z-eicosatetraenoic acid (12-HETE) was stimulated by micromolar concentrations of 13S-hydroperoxy-9Z,11E-octadecadienoic acid, suggesting metabolic control of the 12-lipoxygenase activity via the hydroperoxide tone. Immunohistochemical investigation revealed that the enzyme is mainly located in the squamous epithelium, and is of platelet-type. Significantly lower values for the 12-HETE formation were found in samples from patients with invasive cervical carcinoma, whereas the amount of immunochemically detectable 12-lipoxygenase protein was unaltered. At the same time the expression levels of the bcl-2 gene were enhanced. Thus, it is concluded that during carcinogenesis the hydroperoxide-reducing capacity of the uterine cervix tissue is enhanced, possibly mediated by bcl-2 protein, and in turn metabolically suppresses the 12-lipoxygenase activity. Furthermore, the data suggest an anti-carcinogenic action of 12-lipoxygenase in human cervix, in contrast to its reported pro-carcinogenic action in breast cancer. Int. J. Cancer 82:827–831, 1999. © 1999 Wiley-Liss, Inc.
- Published
- 1999
19. Effect of Acetate and pH on Sunflower Oil Assimilation by Mucor circinelloides f. circinelloides
- Author
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Jacqueline Jeffery, Johan L.F. Kock, Tankred Schewe, Piet J. Botes, A.S. Bareetseng, Santosh Nigam, A. Botha, D.J. Coetzee, and James C. du Preez
- Subjects
chemistry.chemical_classification ,food.ingredient ,biology ,Sunflower oil ,Industrial fermentation ,Metabolism ,biology.organism_classification ,Applied Microbiology and Biotechnology ,Microbiology ,chemistry.chemical_compound ,food ,chemistry ,Biochemistry ,Dry weight ,Mucor circinelloides ,biology.protein ,Food science ,Lipase ,Sodium acetate ,Ecology, Evolution, Behavior and Systematics ,Polyunsaturated fatty acid - Abstract
Summary Mucor circinelloides f. circinelloides CBS 108.16 exhibited poor cell growth and substrate assimilation (32.5% of extracellular lipids) when grown with sunflower oil as sole carbon source. By contrast, in the presence of both sunflower oil (30 g/l) and sodium acetate (10 g/l), nearly complete utilization of both substrates (97.5% of sunflower oil, 100% of sodium acetate) occured and biomass production was increased about five-fold to 23 g dry weight/l after 72 h. A significantly higher content of γ-linolenic acid in the fungal neutral lipid fraction (5.2%) was also observed after 168 h of growth. These different patterns were attributed to the change in pH of the medium during cell growth in the presence and absence of acetate. In the absence of sodium acetate the pH decreased to 2.2, whereas in its presence it increased to about pH 8.0. During metabolism of sunflower oil in the presence of sodium acetate, the percentage of saturated fatty acids in the medium increased, suggesting a higher specificity of the fungal lipase for unsaturated fatty acids. For growth experiments in a small fermenter in the absence of sodium acetate, the gradual pH increase of sodium acetate containing medium was mimicked by an identical programme-controlled pH increase. Similar results as in the presence of sodium acetate were obtained. This observation indicated that the pH increase alone during cultivation was responsible for the increased sunflower oil utilization, biomass and GLA production.
- Published
- 1999
20. Biological dynamics and distribution of 3-hydroxy fatty acids in the yeast Dipodascopsis uninucleata as investigated by immunofluorescence microscopy. Evidence for a putative regulatory role in the sexual reproductive cycle 1
- Author
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J.L.F Kock, Santosh Nigam, Tankred Schewe, P Venter, and D. Linke
- Subjects
biology ,medicine.diagnostic_test ,Biophysics ,Endogeny ,Cell Biology ,Immunofluorescence ,Biochemistry ,Yeast ,Cell wall ,chemistry.chemical_compound ,Biotransformation ,chemistry ,Structural Biology ,Genetics ,biology.protein ,medicine ,Arachidonic acid ,Antibody ,Molecular Biology ,Developmental biology - Abstract
Dipodascopsis uninucleata has been recently shown to produce 3-hydroxy polyenoic fatty acids from several exogenous polyenoic fatty acids. In order to examine whether endogenous 3-hydroxy fatty acids (3-OH-FA) may be implicated in the developmental biology of this yeast, we mapped by immunofluorescence microscopy their occurrence in fixed cells with or without cell walls using an antibody raised against 3R-hydroxy-5Z,8Z,11Z,14Z-eicosatetraenoic acid (3R-HETE), the biotransformation product from arachidonic acid (AA). This antibody turned out to cross-react with other 3-OH-FA. 3-OH-FA were detected in situ in gametangia, asci, as well as between released ascospores, and proved to be associated with the sexual reproductive stage of the life cycle of the yeast. Acetylsalicylic acid (1 mM), which is known to suppress the formation of 3-OH-FA from exogenous polyenoic fatty acids, inhibited the occurrence of immunoreactive material as well as the sexual phase of the life cycle suggesting a prominent regulatory role of 3-OH-FA for the latter.
- Published
- 1998
21. Production of 3R-hydroxy-polyenoic fatty acids by the yeast Dipodascopsis uninucleata
- Author
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R. K. Bhatt, Tankred Schewe, Santosh Nigam, John R. Falck, G. Sravan Kumar, D.J. Coetzee, Pierre Venter, Piet J. Botes, Alfred Botha, and Johan L.F. Kock
- Subjects
chemistry.chemical_classification ,Molecular Structure ,Metabolite ,Linoleic acid ,Organic Chemistry ,Fatty acid ,Stereoisomerism ,Arachidonic Acids ,Cell Biology ,Hydroxylation ,Biochemistry ,Gas Chromatography-Mass Spectrometry ,Yeast ,Oleic acid ,chemistry.chemical_compound ,Linolelaidic acid ,Ascomycota ,chemistry ,Hydroxyeicosatetraenoic Acids ,Fatty Acids, Unsaturated ,Arachidonic acid ,Oxidation-Reduction ,Polyunsaturated fatty acid - Abstract
Various fatty acids were fed to the yeast Dipodascopsis uninucleata UOFS Y 128, and the extracted samples were analyzed for the accumulation of 3-hydroxy metabolites with the help of electron impact gas chromatography-mass spectrometry. Fatty acids containing of 5Z,8Z-diene system (5Z,8Z,11Z-eicosatrienoic, 5Z,8Z,11Z,14Z-eicosatetraenoic, and 5Z,8Z,11Z,14Z,17Z-eicosapentaenoic acids) yielded the corresponding 3-hydroxy-all-Z-eicosapolyenoic acids. Moreover, linoleic acid (9Z,12Z-octadecadienoic acid) and 11Z,14Z,17Z-eicosatrienoic acid were converted to the 3-hydorxylated metabolites of shorter chain length, e,g., 3-hydroxy-5Z,8Z-tetradecadienoic acid and 3-hydroxy-5Z,8Z,11Z-tetradecatrienoic acid, respectively. In contrast, no accumulation of a 3-hydroxy metabolite was observed with oleic acid (9Z-octadecenoic acid), linolelaidic acid (9E,12E-octadecadienoic acid), gamma-linolenic acid (6Z,9Z,12Z-octadecatrienoic acid), and eicosanoic acid as substrate. These findings pinpoint that the 3-hydroxylation of a fatty acid in Dipodascopsis uninucleata requires a 5Z,8Z-diene system either directly or following initial incomplete beta-oxidation. Following analysis of the enantiomer composition, the arachidonic acid metabolite was identified as 3R-hydroxy-5Z,8Z,11Z,14Z-eicosatetraenoic acid, which rules out a normal beta-oxidation as biosynthetic route to this new class of oxylipins.
- Published
- 1997
22. The suppression of 5-lipoxygenation of arachidonic acid in human polymorphonuclear leucocytes by the 15-lipoxygenase product (15S)-hydroxy-(5Z,8Z,11Z,13E)-eicosatetraenoic acid: structure-activity relationship and mechanism of action
- Author
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Peter Ludwig, Tankred Schewe, Karen Petrich, and Hartmut Kühn
- Subjects
Neutrophils ,Stereochemistry ,Eicosatetraenoic acid ,Cell ,Leukotriene B4 ,Biochemistry ,Substrate Specificity ,law.invention ,Structure-Activity Relationship ,chemistry.chemical_compound ,Lipoxygenase ,law ,Hydroxyeicosatetraenoic Acids ,medicine ,Arachidonate 15-Lipoxygenase ,Humans ,Structure–activity relationship ,Lipoxygenase Inhibitors ,Molecular Biology ,Calcimycin ,Arachidonate 5-Lipoxygenase ,Arachidonic Acid ,biology ,Cell Biology ,Recombinant Proteins ,Kinetics ,medicine.anatomical_structure ,Mechanism of action ,chemistry ,Fatty Acids, Unsaturated ,biology.protein ,Recombinant DNA ,Arachidonic acid ,medicine.symptom ,Enantiomer ,Research Article - Abstract
(15S)-Hydroxy-(5Z,8Z,11Z,13E)-eicosatetraenoic acid (15-HETE) suppresses in ionophore-{"type":"entrez-nucleotide","attrs":{"text":"A23187","term_id":"833253","term_text":"A23187"}}A23187-stimulated human polymorphonuclear leucocytes (PMN) the conversion of exogenous arachidonic acid into leukotriene B(4) (LTB4) and (5S)-hydroxy-(6E,8Z,11Z,14Z)-eicosatetraenoic acid (5-HETE). However, contrary to earlier suggestions, 15-HETE is not a genuine 5-lipoxygenase inhibitor under these conditions, but rather suppresses the 5-lipoxygenation of arachidonic acid by switching-over of substrate utilization, as judged from a sizeable formation of labelled (5S,15S)-dihydroxy-(6E,8Z,11Z,13E)-eicosatetr aen oic acid (5,15-diHETE) from 15-[1(-14)C]HETE. Identical results were obtained with human recombinant 5-lipoxygenase. In PMN the formation of 5,15-diHETE is strongly stimulated by either hydroperoxypolyenoic fatty acids or arachidonic acid, suggesting a crucial role of the hydroperoxide tone of the cell. A comparison of a selection of hydroxypolyenoic fatty acids with respect to their capability of suppressing 5-lipoxygenation of arachidonic acid revealed that 15-mono-hydroxyeicosanoids throughout exhibit the highest inhibitory potencies, whereas the other HETEs, 5,15-diHETE as well as octadecanoids, are modest or poor inhibitors. The R and S enantiomers of 15-HETE do not differ from each other, excluding a receptor-like binding of the 15-hydroxy group.
- Published
- 1996
23. Oxygenation of biomembranes by mammalian lipoxygenases: The role of ubiquinone
- Author
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Martina Hellwing, Kerstin Schnurr, Hartmut Kühn, Samuel M. Rapoport, Tankred Schewe, Britta Seidemann, and Peter Jungblut
- Subjects
Erythrocytes ,Free Radicals ,Ubiquinone ,Radical ,Oxidative phosphorylation ,Biochemistry ,Mitochondria, Heart ,Electron Transport ,Superoxide dismutase ,Lipid peroxidation ,chemistry.chemical_compound ,Multienzyme Complexes ,Physiology (medical) ,Animals ,Arachidonate 15-Lipoxygenase ,Vitamin E ,Electrophoresis, Gel, Two-Dimensional ,NADH, NADPH Oxidoreductases ,Submitochondrial particle ,Amino Acids ,Chromatography, High Pressure Liquid ,Heart metabolism ,Mammals ,biology ,Chemistry ,Succinate dehydrogenase ,Cell Membrane ,Keto Acids ,Oxygen ,Succinate Dehydrogenase ,Models, Chemical ,Catalase ,Fatty Acids, Unsaturated ,biology.protein ,Cattle ,Lipid Peroxidation ,Rabbits - Abstract
15-Lipoxygenase is implicated in the selective breakdown of mitochondria during red cell maturation by virtue of its capability of directly oxygenating phospholipids. To address the reason of the selectivity for mitochondria, we studied the reaction of pure rabbit 15-lipoxygenase with beef heart submitochondrial particles in vitro. This reaction is characterised by a loss of polyenoic fatty acids, the formation of phospholipid-bound hydroperoxy- and keto-polyenoic fatty acids, and oxidative modification of membrane proteins. The total oxygen uptake exceeds the formation of oxygenated polyenoic fatty acids several times. The excessive oxygen uptake was not inhibited by 3,5-di-tert-butyl-4-hydroxytoluene or by respiratory inhibitors, but was partly suppressed by superoxide dismutase plus catalase, salicylate, or mannitol. Pentane-extraction of the submitochondrial particles abolished the excessive oxygen uptake, whereas reconstitution with ubiquinone- 50 restored it. A marked excessive oxygen uptake did not occur during the analogous reaction with erythrocyte ghosts. It is proposed that ubiquinone-50 triggers the formation of hydroxyl radicals from 15-lipoxygenase-derived hydroperoxy-lipids via a Fenton-type reaction driven by ubisemiquinone radicals. A new prooxidative function of ubiquinone in the biologically programmed degradation of mitochondria in certain types of cells is proposed.
- Published
- 1996
24. 3,5-Di-t-butyl-4-hydroxytoluene (BHT) and probucol stimulate selectively the reaction of mammalian 15-lipoxygenase with biomembranes
- Author
-
Samuel M. Rapoport, Hartmut Kühn, Kerstin Schnurr, and Tankred Schewe
- Subjects
Linoleic acid ,Biophysics ,Probucol ,Stimulation ,Biochemistry ,Antioxidants ,Mitochondria, Heart ,chemistry.chemical_compound ,Lipoxygenase ,Oxygen Consumption ,Endocrinology ,medicine ,Animals ,Arachidonate 15-Lipoxygenase ,Humans ,Butylated hydroxytoluene ,Phospholipids ,Fluorescent Dyes ,Mammals ,chemistry.chemical_classification ,Quenching (fluorescence) ,biology ,Erythrocyte Membrane ,Fatty acid ,Butylated Hydroxytoluene ,Membrane ,chemistry ,biology.protein ,Cattle ,Rabbits ,medicine.drug - Abstract
The lipophilic antioxidant 3,5-di-t-butyl-4-hydroxytoluene (BHT) and the structurally-related antiatherogenic drug probucol stimulate the oxygenation of mitochondrial membranes and erythrocyte ghosts by the rabbit 15-lipoxygenase as indicated by an increase in oxygen consumption as well as by an enhanced loss of polyenoic fatty acids and by the formation of specific lipoxygenase products in the membrane phospholipids. The oxygenation of linoleic acid, phospholipids and human low-density lipoproteins was not stimulated. With mitochondrial membranes, BHT causes a quenching of the 1-anilino-8-naphthalene sulfonate fluorescence. Thus, it is suggested that the stimulation of membrane oxygenation may be due to structural changes in the membranes leading to a better susceptibility of the polyenoic fatty acid residues towards lipoxygenase attack. Owing to this unexpected effect of the antioxidants, which is not related to their radical-scavenger capacity, care should be taken in interpreting experimental data on effects of BHT and probucol.
- Published
- 1995
25. Strong inhibition of mammalian lipoxygenases by the antiinflammatory seleno-organic compound ebselen in the absence of glutathione
- Author
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Tankred Schewe, Christiane Schewe, and Albrecht Wendel
- Subjects
Azoles ,Lipid Peroxides ,Linoleic acid ,Isoindoles ,Biochemistry ,chemistry.chemical_compound ,Lipoxygenase ,Organoselenium Compounds ,medicine ,Animals ,Humans ,Cyclooxygenase Inhibitors ,Lipoxygenase Inhibitors ,Pharmacology ,chemistry.chemical_classification ,biology ,Ebselen ,Anti-Inflammatory Agents, Non-Steroidal ,Glutathione ,Enzyme ,Linoleic Acids ,chemistry ,Mechanism of action ,Enzyme inhibitor ,Arachidonate 5-lipoxygenase ,biology.protein ,Rabbits ,medicine.symptom - Abstract
Both human recombinant 5-lipoxygenase (EC 1.13.11.34) and 15-lipoxygenase (EC 1.13.11.33, mammalian enzyme) purified from rabbit reticulocytes were inhibited in the absence of glutathione (GSH) by submicromolar concentrations of the seleno-organic compound ebselen. These concentrations were comparable to those of the enzymes. Soybean lipoxygenase-1 (EC 1.13.11.33, plant enzyme) was not inhibited, whereas prostaglandin endoperoxide synthase-1 (EC 1.14.99.1) was inhibited only at much higher concentrations of ebselen ( ic 50 = 37.7 ± 4.3 μM ). The action of ebselen on reticulocyte 15-lipoxygenase (1050 = 0.17 ± 0.01 μM ) was studied in detail. Inhibition occurred instantaneously and appeared to be reversible and was largely abolished by a 20-fold molar excess of GSH over ebselen. In the presence of 1 mM GSH 50% inhibition was observed only at ebselen concentrations as high as 234 ± 27 μM . 13 S -hydroperoxy-9 Z ,11 E -octadecadienoic acid, the lipoxygenase product formed from linoleic acid, augmented the inhibitory effect at low concentrations and caused a partial reversal at high concentrations. A variety of derivatives or structural analogues of ebselen were also tested and proved to be either inactive or weaker inhibitors of 15-lipoxygenase. We have concluded that the potent inhibition of 15-lipoxygenase by ebselen is due neither to GSH peroxidase-like activity nor to lowering of the hydroperoxide tone. The pharmacological implications of these unique characteristics of the action of ebselen on lipoxygenases are then discussed.
- Published
- 1994
26. Specificity of soybean lipoxygenase-1 in hydrated reverse micelles of sodiumbis(2-ethylhexyl)sulfosuccinate (aerosol OT)
- Author
-
Tankred Schewe, Tatyana N. Shkarina, and Hartmut Kühn
- Subjects
chemistry.chemical_classification ,Aqueous solution ,Chromatography ,biology ,Linoleic acid ,Organic Chemistry ,Fast protein liquid chromatography ,Cell Biology ,Biochemistry ,Micelle ,chemistry.chemical_compound ,Lipoxygenase ,Enzyme ,Pulmonary surfactant ,chemistry ,biology.protein ,Arachidonic acid - Abstract
Soybean lipoxygenase-1 (EC 1.13.11.33) was purified by fast protein liquid chromatography on a MONO Q column and studied with respect to the conversion of linoleic and arachidonic acids in reverse micelles of sodiumbis(2-ethylhexyl)sulfosuccinate inn-octane. In this system the specific activities were lower by one order of magnitude than those in the corresponding aqueous system. High-performance liquid chromatography analyses indicated the predominant formation of 13S-hydroperoxy-9Z,11E-octadecadienoic acid (13-HpODE) from linoleic acid and of 15S-hydroperoxy-5Z,8Z,11Z,13E-eicosatetraenoic acid (15) from arachidonic acid both in the aqueous system and in the reverse micelles. After sedimentation of the hydrated reverse micelles by ultracentrifugation, both linoleic acid and 13-HpODE were found to be enriched in the micelles with only small amounts of these compounds present inn-octane. It is proposed that substrates and products of the lipoxygenase reaction are located mainly in the surfactant shell of the hydrated reverse micelles and reach the micelle-entrapped enzyme by diffusion into the aqueous interior space.
- Published
- 1992
27. Dietary Flavanols: Biochemical Basis of Short-Term and Longer-Term Vascular Responses
- Author
-
Yvonne Steffen, Elisabeth Kravets, Helmut Sies, and Tankred Schewe
- Subjects
Communication ,business.industry ,Physiology ,Medicine ,business ,Term (time) - Published
- 2009
28. List of Contributors
- Author
-
FUMIYOSHI ABE, GIOVANNI ADDOLORATO, PETER ALDRED, R. ALLER, PAULO ALMEIDA, JESÚS-ROMÁN MARTÍNEZ ÁLVAREZ, A.V. NIEUW AMERONGEN, MOGENS L. ANDERSEN, HITOSHI AOSHIMA, SAKAE ARIMOTO-KOBAYASHI, ALEMAYEHU ASFAW, ERICA WEINTRAUB AUSTIN, WERNER BACK, SEUNG JOON BAEK, WANDA BAER-DUBOWSKA, AQUILES A. BARROS, HANS BECKER, BAKAN BÉNÉDICTE, STEFANIE BERWANGER, STEFAN BLEICH, JEFFREY B. BLUMBERG, JERINA BOELENS, GREGOR BOHR, MARC BRACKE, H.S. BRAND, M.L. BRUINS, STEFANO BUIATTI, RODRIGO R. CATHARINO, M. LUISA CERVERA, CHUNG-YEN CHEN, QIAO QIAO CHEN, YUONNES CHEN, GIUSEPPE COMI, SONIA CORTACERO-RAMÍREZ, L. DAENEN, L. De COOMAN, D.A. De LUIS, D.P. De SCHUTTER, MAX L. DEINZER, F. DELVAUX, RALF DEMMEL, G. DERDELINCKX, ESTERA SZWAJCER DEY, M.E. DÍAZ-RUBIO, J. RICHARD DICKINSON, MARION DIDIER, FRIEDHELM DIEL, SUSANNE DIEL, REINHARD A. DILLER, ZODWA DLAMINI, AGNIESZKA DOBROWOLSKA-ZACHWIEJA, JEAN-PIERRE DUFOUR, MARCOS N. EBERLIN, KATHARINA E. EFFENBERGER, GRAHAM EYRES, PAOLO FANTOZZI, JUSTIN A. FEGREDO, PETER FEICK, ALBERTO FERNÁNDEZ-GUTIÉRREZ, ISABEL M.P.L.V.O. FERREIRA REQUIMTE, ANNA FERRULLI, MARTA FONTANA, GLEN P. FOX, JOSÉ Da CRUZ FRANCISCO, NORBERT FRANK, ANDREAS FRANKE, GARY FREEMAN BRI, CORAZON FRIAS, SONJA FRÖLICH, DIETMAR FUCHS, ANDRZEJ GAMIAN, S. GARCÍA-FALCÓN, SALVADOR GARRIGUES, ANTONIO GASBARRINI, GIOVANNI GASBARRINI, CLARISSA GERHÄUSER, ANDREAS GERLOFF, ANDREA GHISELLI, A.M. GIL, I. GOÑI, STANISLAVA GORJANOVIĆ, MIGUEL de la GUARDIA, N.P. GUERRA, LUIS F. GUIDO, SANJAY GUPTA, LINDA HELLBORG, GÜNTER HENZE, MARÍA PURIFICACIÓN HERNÁNDEZ-ARTIGA, JAVIER HERNÁNDEZ-BORGES, MARIA HERWALD, THOMAS HILLEMACHER, SHEIKH JULFIKAR HOSSAIN, PAUL HUGHES, STACEY J.T. HUST, KEVIN HUVAERE, GIUSEPPE IACOMINO, EWA IGNATOWICZ, KATSUMI IKEDA, PAVEL JANDERA, KRISTINA JENETT-SIEMS, RAVIN JUGDAOHSINGH, MASATO KAWASAKI, WILLIAM C. KERR, IGOR KHMELINSKII, YOSHINOBU KISO, HIROFUMI KODA, M. KOMAITIS, KEIJI KONDO, VIOLETTA KRAJKA-KUZNIAK, ROBERTO KRATKY, L. DARREN KRUISSELBRINK, ALAN K.H. LAI, M. LEDOCHOWSKI, SEONG-HO LEE, LORENZO LEGGIO, HUI-JING LI, KRIENGSAK LIRDPRAPAMONGKOL, ALEX G. LITTLE, C. LÓPEZ-MACÍAS, SUZANNE LORET, ZACHARENIA LOUKOU, VALENTIN LOZANOV, SOFIE LUST, JEAN ROBERT MABIALA-BABELA, MARISA MANZANO, OMBRETTA MARCONI, PEDRO MARQUES-VIDAL, COLIN R. MARTIN, E. MARTÍNEZ-CARBALLO, ALPHONSE MASSAMBA, HEIDI MAYER, ZUKILE MBITA, ADELE Mc KINNEY, NIALL McCRAE, GARRY MENZ, RACHEL MEYNELL, DOLORES BELLIDO MILLA, STUART R. MILLIGAN, LUIGI MONTANARI, FRANCISCO J. MORALES, YUJI MORIWAKI, KENNETH J. MUKAMAL, RENÉ J.L. MURPHY, MAOURA NANADOUM, MIRELLA NARDINI, FAUSTA NATELLA, SIMÓN NAVARRO, JENNIFER NICOLAI, HAJIME NOZAWA, FRITZ OFFNER, L.M. PASTRANA-CASTRO, DOUGLAS E. PAULL, ANDREA PAVSLER, C. PEHL, JARA PÉREZ-JIMÉNEZ, BRUCE E. PINKLETON, JURE PISKUR, PAWEL POHL, SAM POSSEMIERS, JACQUES POURQUIE, JONATHAN J. POWELL, VICTOR R. PREEDY, C. PROESTOS, ARAM PROKOP, SANDRA RAINIERI, RAJKUMAR RAJENDRAM, BRITTANY B. RAYBURN, WILLIAM F. RAYBURN, HERBERT M. RIEPL, JOSÉ RODRIGUES, MIGUEL ÁNGEL RODRIGUEZ-DELGADO, OLIVER ROSE, NEIL E. ROWLAND, GIAN LUIGI RUSSO, IKUO SAIKI, D. SAISON, SHUSO SAKUMA, HIROAKI SAKURAI, PAT SANDRA, FULGENCIO SAURA-CALIXTO, ALEXANDRA C.H.F. SAWAYA, CRISTINA SCACCINI, H. SCHENNACH, TANKRED SCHEWE, K. SCHROECKSNADEL, CAROLA SCHUBERT, ANTONIO SEGURA-CARRETERO, H. SEIDL, JOSÉ SERRANO, TAKAYUKI SHIBAMOTO, SANJEEV SHUKLA, HELMUT SIES, EWA SIKORSKA, MAREK SIKORSKI, J. SIMAL-GÁNDARA, MANFRED V. SINGER, EDUARDO V. SOARES, RAJAVENTHAN SRIRAJASKANTHAN, KOJI SUZUKI, JISNUSON SVASTI, HANNA SZAEFER, IDOLO TEDESCO, HIROYASU TOBE, DOMENICA TONELLI, A. TORRADO-AGRASAR, FRANCO TUBARO, VICTORIA VALLS-BELLÉS, BARBARA VANHOECKE, GERD VANHOENACKER, E.C.I. VEERMAN, NURIA VELA, H. VERACHTERT, WILLY VERSTRAETE, K.J. VERSTREPEN, ANTONIO LUIS VILLARINO-MARÍN, JOE A. VINSON, GÜNTER VOLLMER, FRANK VRIESEKOOP, CAROLINE WALKER, S. GOYA WANNAMETHEE, JOHANNES WESTENDORF, GRETHE WIBETOE, TOM Van De WIELE, C. WINKLER, HELEN WISEMAN, MAX C.Y. WONG, OWEN L. WOODMAN, SASCHA WUNDERLICH, JIN-WEN XU, HIROAKI YAJIMA, TETSUYA YAMAMOTO, ARUTO YOSHIDA, CHARLES Y.F. YOUNG, PAOLA ZANOLI, MANUELA ZAVATTI, FENG ZHAO, MAŁGORZATA ZIELINSKA-PRZYJEMSKA, OLIVER ZIERAU, and ANASTASIA ZOTOU
- Published
- 2009
29. Epicatechin and Its Role in Protection of LDL and of Vascular Endothelium
- Author
-
Helmut Sies and Tankred Schewe
- Subjects
biology ,Superoxide ,Pharmacology ,Nitric oxide ,chemistry.chemical_compound ,chemistry ,Biochemistry ,In vivo ,Myeloperoxidase ,Blood plasma ,biology.protein ,Peroxynitrite ,Ex vivo ,Lipoprotein - Abstract
(–)-Epicatechin is a dietary polyphenol exerting beneficial effects on the cardiovascular system as judged from epidemiological and clinical studies. Cocoa products, red wine as well as green and black tea are most prominent sources for (–)-epicatechin and related flavan-3-ols. The possible contribution of beer still remains to be substantiated. Comparison of the uptake and elimination kinetics as well as of plasma peak concentrations revealed that among dietary flavan-3-ols, (–)-epicatechin exhibits sufficient bioavailability. The catechol arrangement of the B ring in flavan-3-ols accounts for radical-scavenging, reducing and metal ion-chelating properties in vitro, which may play a protective role in the gastrointestinal tract, whereas for the metabolites in blood plasma these properties are largely lost if this grouping is blocked through glucuronidation and/or methylation. This fact may explain why flavan-3-ols and other flavonoids show strong anti-oxidant activities in vitro such as inhibition of low-density lipoprotein (LDL) oxidation, whereas the corresponding activities in vivo or ex vivo are only moderate. Flavan-3-ols serve anti-oxidant functions in a broader sense suppressing reactions of prooxidant enzymes such as myeloperoxidase, lipoxygenases and NADPH oxidases. High flavan-3-ol intake through a model meal or beverage gives rise to anti-atherosclerotic, anti-inflammatory and anti-platelet activities in vivo as well as improvement of endothelial function of arterial vessels, and lowering of blood pressure. Part of these actions is closely connected with modulation of nitric oxide metabolism of vascular endothelium. In vitro, (–)-epicatechin protects vascular endothelial cells against a number of cytotoxic and proapoptotic actions of oxLDL. Mechanistically, the protective actions involve prevention and scavenging of superoxide anion radical, peroxynitrite and nitrogen dioxide radical, collectively leading to elevation of bioavailability and bioactivity of nitric oxide and, in turn, to improvement of endothelial function.
- Published
- 2009
30. Mono-O-methylated flavanols and other flavonoids as inhibitors of endothelial NADPH oxidase
- Author
-
Yvonne Steffen, Claudia Gruber, Helmut Sies, and Tankred Schewe
- Subjects
Umbilical Veins ,Endothelium ,Flavonols ,Biophysics ,medicine.disease_cause ,Biochemistry ,Umbilical vein ,Catechin ,Nitric oxide ,chemistry.chemical_compound ,Structure-Activity Relationship ,medicine ,Biflavonoids ,Humans ,Proanthocyanidins ,Enzyme Inhibitors ,Molecular Biology ,Procyanidin B2 ,Cells, Cultured ,Flavonoids ,NADPH oxidase ,biology ,NADPH Oxidases ,Flavones ,medicine.anatomical_structure ,chemistry ,Drug Design ,Microsome ,biology.protein ,Microsomes, Liver ,Quercetin ,Endothelium, Vascular ,Intracellular ,Oxidative stress - Abstract
The dietary flavan-3-ol (-)-epicatechin improves the bioactivity of nitric oxide in arterial vessels in vivo. Moreover, it effectively protects cultured vascular endothelial cells from signs of oxidative stress and elevates intracellular nitric oxide in vitro. We addressed the effects of (-)-epicatechin, its metabolic conversion products and structurally related compounds on NADPH oxidase activity in intact human umbilical vein endothelial cells (HUVEC) and in cell lysates. (-)-Epicatechin proved to be an O2*(-)-scavenger but did not inhibit NADPH oxidase activity, whereas the converse pattern was observed for the metabolites 3'- and 4'-O-methyl epicatechin. The dimer procyanidin B2 and (-)-epicatechin glucuronide were O2*(-)-scavengers and inhibited NADPH oxidase. Analysis of structure-activity relations with 45 compounds suggests an apocynin-like mode of NADPH oxidase inhibition. Notably, HUVEC converted (-)-epicatechin to NADPH oxidase-inhibitory methyl ethers. These data identify endothelial NADPH oxidase as candidate target of dietary flavonoids and particularly of their metabolites.
- Published
- 2007
31. Nitrite, a naturally occurring precursor of nitric oxide that acts like a 'prodrug'
- Author
-
Klaus-Dietrich Kröncke, Christoph V. Suschek, Helmut Sies, and Tankred Schewe
- Subjects
Saliva ,Clinical Biochemistry ,Vasodilation ,Nitric Oxide ,Biochemistry ,Nitric oxide ,chemistry.chemical_compound ,medicine ,Animals ,Humans ,Prodrugs ,Nitrite ,Molecular Biology ,Cell damage ,Nitrites ,Skin ,chemistry.chemical_classification ,Nitrates ,integumentary system ,Hypoxia (medical) ,Prodrug ,medicine.disease ,Enzyme ,chemistry ,medicine.symptom - Abstract
There are enzymatic and non-enzymatic mechanisms that generate NO* from nitrite in blood, stomach, saliva, urine and skin. In blood vessels, nitrite-derived NO* can provide protection via compensatory vasodilation during hypoxia, and in various body fluids it may have antibacterial activity. In the skin, nitrite-derived NO* may contribute to skin tanning, as well as to protection against UV-induced cell damage. Current knowledge on nitrite acting like an NO* 'prodrug' is presented, emphasizing the role of nitrite in skin.
- Published
- 2006
32. Cytotoxicity of myeloperoxidase/nitrite-oxidized low-density lipoprotein toward endothelial cells is due to a high 7beta-hydroxycholesterol to 7-ketocholesterol ratio
- Author
-
Helmut Sies, Yvonne Steffen, Daniela Peter, Tankred Schewe, and Ingrid Wiswedel
- Subjects
Umbilical Veins ,Oxysterol ,Cell Survival ,medicine.disease_cause ,Biochemistry ,Catechin ,chemistry.chemical_compound ,Physiology (medical) ,polycyclic compounds ,medicine ,Humans ,Nitrite ,Cytotoxicity ,Ketocholesterols ,Cells, Cultured ,Nitrites ,Peroxidase ,NADPH oxidase ,biology ,Endothelial Cells ,Molecular biology ,Hydroxycholesterols ,Lipoproteins, LDL ,chemistry ,Myeloperoxidase ,Apocynin ,biology.protein ,lipids (amino acids, peptides, and proteins) ,Endothelium, Vascular ,Oxidation-Reduction ,Oxidative stress ,Lipoprotein - Abstract
Oxygenated cholesterols (oxysterols) formed during oxidation of low-density lipoprotein (LDL) are associated with endothelial dysfunction and atherogenesis. We compared the profile of oxysterols in modified human LDL obtained on reaction with myeloperoxidase/H2O2 plus nitrite (MPO/H2O2/nitrite-oxLDL) with that on Cu2+ -catalyzed oxidation. The 7beta-hydroxycholesterol/7-ketocholesterol ratio was markedly higher in MPO/H2O2/nitrite-oxLDL than in Cu2+ -oxidized LDL (7.9 +/- 3.0 versus 0.94 +/- 0.10). Like MPO/H2O2/nitrite-oxLDL, 7beta-hydroxycholesterol was cytotoxic toward endothelial cells through eliciting oxidative stress. Cytotoxicity was accompanied by DNA fragmentation and was prevented by the NADPH oxidase inhibitor apocynin, suggesting stimulation of NADPH oxidase-mediated O2-* formation. 7-Ketocholesterol was only cytotoxic when added alone, whereas a 1:1-mixture with 7beta-hydroxycholesterol surprisingly was noncytotoxic. We conclude from our data that (i) 7beta-hydroxycholesterol is a pivotal cytotoxic component of oxidized LDL, (ii) 7-ketocholesterol protects against 7beta-hydroxycholesterol in oxysterol mixtures or oxLDL, (iii) the 7beta-hydroxycholesterol/7-ketocholesterol ratio is a crucial determinant for cytotoxicity of oxidized LDL species and oxysterol mixtures, and (iv) the low share of 7-ketocholesterol explains the higher cytotoxicity of MPO/H2O2/nitrite-oxLDL than other forms of oxidized LDL. The dietary polyphenol (-)-epicatechin inhibited not only formation but also cytotoxic actions of both oxLDL and oxysterols.
- Published
- 2006
33. OxLDL signaling in endothelial cells and counteracting effects of (-)-epicatechin
- Author
-
Yvonne Steffen, Helmut Sies, and Tankred Schewe
- Published
- 2006
34. Oxidative modification of low-density lipoprotein: lipid peroxidation by myeloperoxidase in the presence of nitrite
- Author
-
Inthanongsack Prakosay, Tilo Kraemer, Rahul A Date, Helmut Sies, and Tankred Schewe
- Subjects
Antioxidant ,medicine.medical_treatment ,Clinical Biochemistry ,Oxidative phosphorylation ,Biochemistry ,Lipid peroxidation ,chemistry.chemical_compound ,medicine ,Humans ,Nitrite ,Molecular Biology ,Nitrites ,Peroxidase ,Flavonoids ,biology ,Dose-Response Relationship, Drug ,Lipoproteins, LDL ,Oxidative Stress ,chemistry ,Low-density lipoprotein ,Myeloperoxidase ,biology.protein ,lipids (amino acids, peptides, and proteins) ,Lipid Peroxidation ,Peroxynitrite ,Lipoprotein - Abstract
Oxidative modification of low-density lipoprotein (LDL) is a pivotal process in early atherogenesis and can be brought about by myeloperoxidase (MPO), which is capable of reacting with nitrite, a NO metabolite. We studied MPO-mediated formation of conjugated dienes in isolated human LDL in dependence on the concentrations of nitrite and chloride. This reaction was strongly stimulated by low concentrations (5-50 microM) of nitrite which corresponds to the reported concentration in the arterial vessel wall. Under these conditions no protein tyrosine nitration occurred; this reaction required much higher nitrite concentrations (100 microM-1 mM). Chloride neither supported lipid peroxidation alone nor was its presence mandatory for the effect of nitrite. We propose a prominent role of lipid peroxidation for the proatherogenic action of the MPO/nitrite system, whereas peroxynitrite may be competent for protein tyrosine nitration of LDL. Monomeric and oligomeric flavan-3-ols present in cocoa products effectively counteracted, at micromolar concentrations, the MPO/nitrite-mediated lipid peroxidation of LDL. Flavan-3-ols also suppressed protein tyrosine nitration induced by MPO/nitrite or peroxynitrite as well as Cu2+-mediated lipid peroxidation of LDL. This multi-site protection by (-)-epicatechin or other flavan-3-ols against proatherogenic modification of LDL may contribute to the purported beneficial effects of dietary flavan-3-ols for the cardiovascular system.
- Published
- 2004
35. Inhibition of 15-lipoxygenases by flavonoids: structure-activity relations and mode of action
- Author
-
Helmut Sies, Christian D. Sadik, and Tankred Schewe
- Subjects
Stereochemistry ,Biochemistry ,Flavones ,chemistry.chemical_compound ,Structure-Activity Relationship ,medicine ,Animals ,Lipoxygenase Inhibitors ,Enzyme Inhibitors ,Mode of action ,Pharmacology ,chemistry.chemical_classification ,Flavonoids ,biology ,Baicalein ,Kinetics ,chemistry ,Mechanism of action ,Enzyme inhibitor ,biology.protein ,Quercetin ,Rabbits ,medicine.symptom ,Luteolin ,Fisetin - Abstract
We have recently reported that flavonoids of cocoa inhibit the mammalian 15-lipoxygenase-1-a catalyst of enzymatic lipid peroxidation. To elucidate the structure-activity relationship of the inhibitory effect, we investigated the effects of 18 selected flavonoids of variable structure on pure rabbit reticulocyte and soybean 15-lipoxygenases using linoleic acid as substrate. Moreover, the inhibition by quercetin was studied in detail to gain insight into the mode of action. Quercetin was found to modulate the time-course of the reaction of both lipoxygenases by three distinct effects: (i) prolongation of the lag period, (ii) rapid decrease in the initial rate after the lag phase was overcome, (iii) time-dependent inactivation of the enzyme during reaction but not in the absence of substrate. A comparison of the IC(50) for the rapid inhibition of rabbit reticulocyte 15-lipoxygenase-1 revealed that (i) the presence of a hydroxyl group in the flavonoid molecule is not essential, (ii) a catechol arrangement reinforces the inhibitory effect, (iii) in the presence of a catechol arrangement the inhibitory potency inversely correlates with the number of hydroxyl groups, (iv) a 2,3-double bond in the C ring strengthens the inhibitory effect. The flavone luteolin turned out to be the most potent inhibitor of the mammalian enzyme with an IC(50) of 0.6 microM followed by baicalein (1 microM) and fisetin (1.5 microM).
- Published
- 2003
36. Myeloperoxidase/nitrite-mediated lipid peroxidation of low-density lipoprotein as modulated by flavonoids
- Author
-
Tankred Schewe, Vladimir A. Kostyuk, Helmut Sies, and Tilo Kraemer
- Subjects
Oxidized LDL ,Rutin ,Biophysics ,Biochemistry ,Catechin ,Lipid peroxidation ,chemistry.chemical_compound ,Structural Biology ,Genetics ,Taxifolin ,Humans ,heterocyclic compounds ,Nitrite ,Molecular Biology ,Nitrites ,Peroxidase ,Flavonoids ,biology ,food and beverages ,Reactive nitrogen species ,Cell Biology ,Hydrogen Peroxide ,Atherosclerosis ,Lipoproteins, LDL ,Kinetics ,chemistry ,Low-density lipoprotein ,Myeloperoxidase ,biology.protein ,lipids (amino acids, peptides, and proteins) ,Flavanol ,Quercetin ,Lipid Peroxidation ,Luteolin ,Flavonol - Abstract
In the presence of a H2O2-generating system, myeloperoxidase (MPO) caused conjugated diene formation in low-density lipoprotein (LDL), indicating lipid peroxidation which was dependent on nitrite but not on chloride. The oxidation of LDL was inhibited by micromolar concentrations of flavonoids such as (−)-epicatechin, quercetin, rutin, taxifolin and luteolin, presumably via scavenging of the MPO-derived NO2 radical. The flavonoids served as substrates of MPO leading to products with distinct absorbance spectra. The MPO-catalyzed oxidation of flavonoids was accelerated in the presence of nitrite.
- Published
- 2003
37. Polyphenols of cocoa: inhibition of mammalian 15-lipoxygenase
- Author
-
Tanihiro Yoshimoto, Hartmut Kühn, Helmut Sies, Christian D. Sadik, Lars-Oliver Klotz, and Tankred Schewe
- Subjects
Reticulocytes ,Polymers ,Swine ,Clinical Biochemistry ,Prostacyclin ,Epigallocatechin gallate ,Transfection ,Biochemistry ,Catechin ,chemistry.chemical_compound ,Lipoxygenase ,Mice ,Phenols ,Eicosanoic Acids ,Hydroxyeicosatetraenoic Acids ,medicine ,Animals ,Arachidonate 15-Lipoxygenase ,Humans ,12-Hydroxy-5,8,10,14-eicosatetraenoic Acid ,Lipoxygenase Inhibitors ,Enzyme Inhibitors ,Molecular Biology ,Cells, Cultured ,Flavonoids ,Leukotriene ,Cacao ,biology ,Plant Extracts ,Macrophages ,Lipoproteins, LDL ,Epicatechin gallate ,chemistry ,Proanthocyanidin ,Polyphenol ,biology.protein ,Arachidonic acid ,Rabbits ,Soybeans ,medicine.drug - Abstract
Some cocoas and chocolates are rich in (-)-epicatechin and its related oligomers, the procyanidins. Fractions of these compounds, isolated from the seeds of Theobroma cacao, caused dose-dependent inhibition of isolated rabbit 15-lipoxygenase-1 with the larger oligomers being more active; the decamer fraction revealed an IC50 of 0.8 microM. Among the monomeric flavanols, epigallocatechin gallate (IC50 = 4 microM) and epicatechin gallate (5 microM) were more potent than (-)-epicatechin (IC50 = 60 microM). (-)-Epicatechin and procyanidin nonamer also inhibited the formation of 15-hydroxy-eicosatetraenoic acid from arachidonic acid in rabbit smooth muscle cells transfected with human 15-lipoxygenase-1. In contrast, inhibition of the lipoxygenase pathway in J774A.1 cells transfected with porcine leukocyte-type 12-lipoxygenase (another representative of the 12/15-lipoxygenase family) was only observed upon sonication of the cells, suggesting a membrane barrier for flavanols in these cells. Moreover, epicatechin (IC50 approx. 15 microM) and the procyanidin decamer inhibited recombinant human platelet 12-lipoxygenase. These observations suggest general lipoxygenase-inhibitory potency of flavanols and procyanidins that may contribute to their putative beneficial effects on the cardiovascular system in man. Thus, they may provide a plausible explanation for recent literature reports indicating that procyanidins decrease the leukotriene/prostacyclin ratio in humans and human aortic endothelial cells.
- Published
- 2002
38. 15-Lipoxygenase-1: A Prooxidant Enzyme
- Author
-
Tankred Schewe
- Subjects
Reticulocytes ,Clinical Biochemistry ,Oxidative phosphorylation ,medicine.disease_cause ,Biochemistry ,Catalysis ,Lipid peroxidation ,chemistry.chemical_compound ,Lipoxygenase ,medicine ,Animals ,Arachidonate 15-Lipoxygenase ,Humans ,Molecular Biology ,chemistry.chemical_classification ,biology ,Cholesterol ,Oxidants ,Isoenzymes ,Enzyme ,chemistry ,biology.protein ,Lipid Peroxidation ,Intracellular ,Oxidative stress ,Lipoprotein - Abstract
Human and rabbit reticulocyte 15-lipoxygenase (15-lipoxygenase-1) and the leukocyte-type 12-lipoxygenases (12/15-lipoxygenases) of pig, beef, mouse and rat constitute a particular subfamily of mammalian lipoxygenases (reticulocyte-type lipoxygenases) with unique properties and functions. They catalyze enzymatic lipid peroxidation in complex biological structures via direct dioxygenation of phospholipids and cholesterol esters of biomembranes and plasma lipoproteins. Moreover, they are a source of free radicals initiating non-enzymatic lipid peroxidation and other oxidative processes. Expression and activity of reticulocyte-type lipoxygenases are highly regulated. Moreover, the susceptibility of intracellular membranes toward these lipoxygenases is controlled and may be increased together with lipoxygenase activity under conditions of oxidative stress. Thus, oxidative stress may favor a concerted package of lipoxygenase-mediated enzymatic and non-enzymatic lipid peroxidation and co-oxidative processes. Reaction of reticulocyte-type lipoxygenases with low-density lipoprotein renders the latter atherogenic and appears to be involved in the formation of atherosclerotic lesions.
- Published
- 2002
39. Novel Aspects Related to Biosynthesis and Biological Actions of Hepoxilins: Interrelationship with Phospholipid Hydroperoxide Glutathione Peroxidase (PHGPx)
- Author
-
Pattabhiraman Shankaranarayanan, Santosh Nigam, Mark Sutherland, Helmar Waiczies, and Tankred Schewe
- Subjects
chemistry.chemical_classification ,biology ,Eicosanoid metabolism ,Glutathione peroxidase ,Glutathione ,Metabolism ,GPX4 ,chemistry.chemical_compound ,Lipoxygenase ,Biochemistry ,chemistry ,biology.protein ,Phospholipid-hydroperoxide glutathione peroxidase ,Peroxidase - Abstract
The 12-lipoxygenase (12-LOX) pathway of AA metabolism in platelets and other cells is bifurcated at the level of the oxygenated product 12-hydroperoxyeicosatetranoic acid (12-HPETE) into a reduction route leading to 12-HETE as a primary product and an isomerization route forming hepoxilins (HXA3 and HXB3) as major products. Both 12-HETE and hepoxilins are biologically active eicosanoids and possess a myriad of biological actions [1,2 and references therein]. Glutathione peroxidases, which are key players in the defense of tissues and cells against the oxidative damage [3], are also intimately involved in the regulation of AA metabolism [4–7]. This role has been attributed to glutathione peroxidases for following reasons: (a) they reduce the hydroperoxyfatty acids formed by lipoxygenases or cyclooxygenases to corresponding alcohols, (b) they regulate the hydroperoxide tone of the system, which is contributed largely by the hydroperoxyfatty acids. It is known that a low hydroperoxide tone functions as a stimulus for lipoxygenase and cyclooxygenase reactions but a higher hydroperoxide tone causes suicide inactivation of dioxygenases and of substrates for hydroperoxidase reaction [8]. Four different glutathione peroxidases, which are characterized by the selenocysteine group at the active site, are known up until now [3]: cytosolic glutathione peroxidase (GPx-1), PHGPx (GPx4), plasma GPx (pGPx), and gastrointestinal GPx (GIGPx). But, only GPx-1 and PHGPx play a significant role in the intracellular metabolism of eicosanoids. Several studies in the past dealing with the role of glutathione peroxidases in the eicosanoid metabolism of platelets considered predominantly GPX-1 as the sole enzyme. To our knowledge no evidence for the coeval occurrence of PHGPx in platelets has been demonstrated so far. The present study provides an evidence for the presence of PHGPx in platelets. Its pivotal role in the regulation of 12-LOX pathway has been demonstrated by the inactivation of glutathione peroxidases by iodoacetate without any inactivation of 12-LOX [9] as well as in selenium-deficient cells.
- Published
- 2001
40. Oxylipin formation in fungi: biotransformation of arachidonic acid to 3-hydroxy-5,8-tetradecadienoic acid by Mucor genevensis
- Author
-
D.J. Coetzee, Johan L.F. Kock, Tankred Schewe, Alfred Botha, Santosh Nigam, Piet J. Botes, and Carolina H. Pohl
- Subjects
Chromatography ,Arachidonic Acid ,Linoleic acid ,Metabolite ,Biophysics ,Cell Biology ,Oxylipin ,Biochemistry ,Yeast ,Mass Spectrometry ,Linoleic Acid ,chemistry.chemical_compound ,chemistry ,Biotransformation ,Mucor ,Hydroxyeicosatetraenoic Acids ,Fatty Acids, Unsaturated ,Arachidonic acid ,Gas chromatography ,Hydroxy Acids ,Molecular Biology ,Soil microbiology ,Soil Microbiology - Abstract
The soil fungus Mucor genevensis was shown to convert exogenous arachidonic acid to the oxylipin 3-hydroxy-5Z,8Z-tetradecadienoic acid (3-HTDE) as determined by gas chromatography/mass spectrometry. This metabolite was only found in the aqueous supernatant together with free linoleic acid, but not in the final fungal biomass. In contrast, the corresponding primary arachidonic acid metabolite (3R)-hydroxy-(5Z,8Z,11Z,14Z)-eicosatetraenoic acid (3-HETE), which has been earlier shown to be produced by the yeast Dipodascopsis uninucleata, could not be detected. These observations may be plausibly explained by a retroconversion by M. genevensis of arachidonic acid to linoleic acid before the latter is metabolised to 3-HTDE.
- Published
- 1999
41. (3R)-Hydroxy-Oxylipins—A Novel Family of Oxygenated Polyenoic Fatty Acids of Fungal Origin
- Author
-
Santosh Nigam, J. Lodewyk F. Kock, and Tankred Schewe
- Subjects
chemistry.chemical_classification ,chemistry.chemical_compound ,chemistry ,Biochemistry ,Arachidonic acid ,Fatty acid derivatives ,Yeast ,Salicylic acid ,Eicosatrienoic Acid ,Polyunsaturated fatty acid ,Sexual reproduction - Abstract
Certain fungi and yeasts are capable of producing hydroxy fatty acids or other oxygenated fatty acid derivatives which are collectively named oxylipinds, and which apparently play a predominant role in the vegetative growth and sexual reproduction of the yeast (Kurtzman et al., 1974; Herman and Herman, 1985; Kerwin et al., 1986; Losel, 1988; Rattray, 1988; Van Dyk et al., 1991; Brodowski and Oliw, 1992; Jensen et al., 1992; Van der Berg, 1993; Van Dyk et al, 1994; Venter et al., 1997; Kock et al., 1998; Herman, 1998).
- Published
- 1999
42. Introductory Remarks
- Author
-
Tankred Schewe, Cecil R. Pace-Asciak, and Santosh Nigam
- Subjects
Lipoxygenase ,biology ,Chemistry ,biology.protein ,Classics - Published
- 1999
43. Production of 3-Hydroxy Fatty Acids by the Yeast Dipodascopsis Uninucleata. Biological Implications
- Author
-
J. Lodewyk F. Kock, Alfred Botha, Tankred Schewe, D. P. Smith, Pieter W. J. van Wyk, Santosh Nigam, D.J. Coetzee, and Pierre Venter
- Subjects
chemistry.chemical_compound ,chemistry ,Biochemistry ,Linoleic acid ,Microorganism ,Hydroxy fatty acid ,Arachidonic acid ,Immunofluorescence Microscopy ,Dipodascopsis uninucleata ,Yeast - Abstract
Hydroxy fatty acids (OH-FAs) including 3-OH-FA are widely distributed in microorganisms (Schweizer, 1989). Thus, the production of 3-OH-FA by fungi has been reported in various studies (Van Dyk et al., 1994). Using thin-layer chromatography, autoradiography, 1H-NMR as well as gas chromatography-mass spectrometry, we isolated and identified the novel 3-hydroxyeicosanoid (3R)-hydroxy-(5Z,8Z,llZ,14Z)-eicosatetraenoic acid (3-HETE) from the yeast Dipodascopsis uninucleata following incubation with arachidonic acid (Van Dyk et al., 1991; Venter et al., 1997). This compound now turned out to be the prototype of the new family of fungal unsaturated bioactive (3R)-hydroxy-oxylipins (see Nigam et al., this volume).
- Published
- 1999
44. Effect of 15-Hete on the 5-Lipoxygenase Pathway in Neutrophils
- Author
-
Santosh Nigam, Tankred Schewe, Hartmut Kühn, Karen Petrich, and Peter Ludwig
- Subjects
Lipoxygenase ,chemistry.chemical_compound ,biology ,Biochemistry ,Chemistry ,Eicosatetraenoic acid ,Arachidonate 5-lipoxygenase ,biology.protein ,Structure–activity relationship ,Substrate (chemistry) ,Arachidonic acid ,Mode of action ,Proinflammatory cytokine - Abstract
In 1976 Borgeat, Hamberg and Samuelsson were the first who clearly demonstrated the presence of the 5-lipoxygenase pathway in leukocytes1. This discovery was an important milestone in the elucidation of leukotriene synthesis. Four years later, in 1980, Van- derhoek, Bryant and Bailey reported on the inhibition of the 5-lipoxygenase pathway of arachidonic acid metabolism of human neutrophil leukocytes by 15-HETE which is in turn a product of the 15-lipoxygenase pathway2. Inasmuch as we meanwhile know that neutro-phils contain both 5- and 15 lipoxygenase activities, it has become a very attractive hypothesis to assume a mutual regulatory relationship between the two lipoxygenase pathways in such a way that the proinflammatory 5-lipoxygenase pathway is controlled by a predominately antiinflammatory 15-lipoxygenase pathway3. The original observation of Vanderhoek and coworkers has been later confirmed by a number of investigators2-10, however, no studies were performed to elucidate the precise mode of action of the inhibitory effect of 15-HETE. The aim of our study was to fill this gap.
- Published
- 1999
45. Is Lipoxygenation of Pathogen-Derived Arachidonic Acid Involved in Plant Protection?
- Author
-
Tankred Schewe and Santosh Nigam
- Subjects
biology ,Jasmonic acid ,fungi ,food and beverages ,Fungal pathogen ,biology.organism_classification ,Microbiology ,chemistry.chemical_compound ,Lipoxygenase ,chemistry ,Algae ,biology.protein ,Arachidonic acid ,Enzyme inducer ,Pathogen - Abstract
In the plant kingdom the occurrence of arachidonic acid and, therefore, of eicosanoids appears to be restricted to fungi and related organisms, algae, mosses and ferns. In higher plants, however, lipoxygenases capable of oxygenating arachidonic acid are widely distributed. At first sight, the reactions of higher plant lipoxygenases with arachidonic acid seem to be only of academic interest. However, under special circumstances a higher plant lipoxygenase is capable of converting arachidonic acid of fungal origin. Such a situation is given by the infection of the plant by a fungal pathogen. Some aspects of the putative role of these interactions will be discussed in this article. For a detailed survey on the present knowledge of the role of lipoxygenase in plant resistance to infection the reader is referred to the recent paper by Slusarenko.1
- Published
- 1997
46. The selenoenzyme phospholipid hydroperoxide glutathione peroxidase controls the activity of the 15-lipoxygenase with complex substrates and preserves the specificity of the oxygenation products
- Author
-
Fulvio Ursini, Jutta Belkner, Hartmut Kühn, Kerstin Schnurr, and Tankred Schewe
- Subjects
GPX1 ,Lipid Peroxides ,Reticulocytes ,Time Factors ,GPX3 ,Submitochondrial Particles ,Biochemistry ,Mitochondria, Heart ,Substrate Specificity ,chemistry.chemical_compound ,Lipoxygenase ,Animals ,Arachidonate 15-Lipoxygenase ,Humans ,Submitochondrial particle ,Phospholipid-hydroperoxide glutathione peroxidase ,Molecular Biology ,chemistry.chemical_classification ,Mammals ,Glutathione Peroxidase ,biology ,Glutathione peroxidase ,Substrate (chemistry) ,Cell Biology ,Intracellular Membranes ,Phospholipid Hydroperoxide Glutathione Peroxidase ,Lipoproteins, LDL ,Kinetics ,chemistry ,Low-density lipoprotein ,Alcohols ,biology.protein ,lipids (amino acids, peptides, and proteins) ,Cattle ,Rabbits ,Oxidation-Reduction - Abstract
Mammalian 15-lipoxygenases have been suggested to be involved in cell differentiation and atherogenesis because of their capability of oxygenating polyenoic fatty acids esterified to biomembranes and lipoproteins. We investigated the interaction of the lipid-peroxidizing 15-lipoxygenase and the hydroperoxy lipid-reducing phospholipid hydroperoxide glutathione peroxidase during their reaction with biomembranes and lipoproteins and obtained the following results. 1) Lipoxygenase treatment of submitochondrial membranes led to the formation of hydroperoxyphosphatidylethanolamine and hydroperoxyphosphatidylcholine as indicated by high performance liquid chromatography with chemiluminescence detection. In 15-lipoxygenase-treated low density lipoprotein cholesteryl hydroperoxylinoleate was the major oxygenation product. 2) Phospholipid hydroperoxide glutathione peroxidase was capable of reducing the hydroperoxy lipids formed by the 15-lipoxygenase to their corresponding alcohols. 3) Preincubation of low density lipoprotein and submitochondrial membranes with the phospholipid hydroperoxide glutathione peroxidase completely prevented the lipoxygenase reaction. However, addition of exogenous hydroperoxy lipids restored the oxygenase activity. 4) Short-term incubations of the complex substrates with the 15-lipoxygenase led to a specific pattern of oxidation products which was rendered more unspecific at long-term incubation or at high substrate concentrations. If the phosholipid hydroperoxide glutathione peroxidase was present during the reaction, the specific product pattern was preserved. These data indicate that the phospholipid hydroperoxide glutathione peroxidase is capable of reducing hydroperoxy ester lipids formed by a 15-lipoxygenase, and that it may down-regulate the 15-lipoxygenase pathways in mammalian cells. The specificity of 15-lipoxygenase-derived hydroperoxy lipids depends on their immediate reduction to the corresponding alcohols preventing postcatalytic isomerization.
- Published
- 1996
47. Inhibition of eicosanoid formation in human polymorphonuclear leukocytes by high concentrations of magnesium ions
- Author
-
Peter Ludwig, Karen Petrich, Wolfgang Diezel, and Tankred Schewe
- Subjects
Leukotriene B4 ,Neutrophils ,In Vitro Techniques ,Biochemistry ,chemistry.chemical_compound ,Hydroxyeicosatetraenoic Acids ,Humans ,Magnesium ,Lipoxygenase Inhibitors ,Mode of action ,IC50 ,Magnesium ion ,Calcimycin ,Chromatography, High Pressure Liquid ,Arachidonate 5-Lipoxygenase ,Arachidonic Acid ,Ionophores ,Eicosanoid metabolism ,Chemistry ,Fatty Acids ,Recombinant Proteins ,N-Formylmethionine Leucyl-Phenylalanine ,Eicosanoid ,Liberation ,Eicosanoids ,Arachidonic acid ,Calcium - Abstract
The cutaneous antiinflammatory action of Dead-Sea brine is thought to be due to magnesium ions. To elucidate their mode of action, we studied the influence of isotonic solutions containing high concentrations of Mg2+ (up to 115mM) on the formation of 5-lipoxygenase-derived eicosanoids in human polymorphonuclear leukocytes. The cells were stimulated by either ionophore A23187 or the chemotactic peptide N-formyl-L-methionyl-L-leucyl-L-phenylalanine. We observed a pronounced inhibition of the formation of leukotriene B4 and 5-hydroxyeicosatetraenoic acid from either added [1-14C] or endogenously liberated arachidonic acid. In the latter case, the sum of arachidonic acid and its oxygenation products was also markedly diminished. The inhibitory effects of Mg2+ depended in a reciprocal manner on the concentration of Ca2+ in the incubation medium. An unspecific damage to cells as reason for the inhibitory effects was excluded. Human recombinant 5-lipoxygenase was also inhibited by Mg2+ in the same concentration range (IC50 16 mM). These data suggest that high concentrations of Mg2+ inhibit the eicosanoid metabolism both at the level of the liberation of arachidonic acid and by direct inhibition of the 5-lipoxygenase enzyme.
- Published
- 1995
48. Molecular actions of ebselen--an antiinflammatory antioxidant
- Author
-
Tankred Schewe
- Subjects
Azoles ,Antioxidant ,medicine.medical_treatment ,Isoindoles ,Antioxidants ,Lipid peroxidation ,Lipoxygenase ,chemistry.chemical_compound ,Organoselenium Compounds ,medicine ,Animals ,Phospholipid-hydroperoxide glutathione peroxidase ,Enzyme Inhibitors ,Protein kinase C ,Pharmacology ,chemistry.chemical_classification ,Inflammation ,Oxidase test ,biology ,Molecular Structure ,Chemistry ,Ebselen ,Enzyme ,Biochemistry ,biology.protein ,Lipid Peroxidation - Abstract
1. 1. Ebselen (2-phenyl-1,2-benzisoselenazol-3(2H)-one) is a non-toxic seleno-organic drug with antiinflammatory, antiatherosclerotic and cytoprotective properties. 2. 2. Ebselen and some of its metabolites are effective reductants of hydroperoxides including those arising in biomembranes and lipoproteins. 3. 3. By reactions with hydroperoxides and thiols several interconversion cycles are formed which include ebselen metabolites with varying oxidation number of the selenium. 4. 4. In the presence of thiols ebselen mimics the catalytic activities of phospholipid hydroperoxide glutathione peroxidase. 5. 5. Ebselen inhibits at low concentrations a number of enzymes involved in inflammation such as lipoxygenases, NO synthases, NADPH, oxidase, protein kinase C and H+/K+-ATPase. The inhibitions are manifested on the cellular level and may contribute to the antiinflammatory potential of ebselen.
- Published
- 1995
49. Vascular Effects of Cocoa Rich in Flavan-3-ols
- Author
-
Andre Dejam, Petra Kleinbongard, Christian Heiss, Helmut Sies, Malte Kelm, and Tankred Schewe
- Subjects
chemistry.chemical_compound ,medicine.anatomical_structure ,chemistry ,Endothelium ,Flavan ,business.industry ,medicine ,General Medicine ,Pharmacology ,Nitric oxide metabolism ,business - Published
- 2003
50. On the reaction specificity of the lipoxygenase from tomato fruits
- Author
-
Dugeriyn Regdel, Tankred Schewe, and Hartmut Kühn
- Subjects
Linoleic acid ,Lipoxygenase ,Biophysics ,Biochemistry ,Mitochondria, Heart ,Substrate Specificity ,Linoleic Acid ,chemistry.chemical_compound ,Endocrinology ,Phosphatidylcholine ,Animals ,Ammonium sulfate precipitation ,chemistry.chemical_classification ,Chromatography ,Arachidonic Acid ,biology ,food and beverages ,alpha-Linolenic Acid ,Fast protein liquid chromatography ,Enzyme ,chemistry ,Linoleic Acids ,Fruit ,biology.protein ,Arachidonic acid ,Cattle ,Polyunsaturated fatty acid - Abstract
A lipoxygenase was purified 300-fold from a homogenate supernatant of ripe tomato fruits by fractionated ammonium sulfate precipitation and anion exchange fast protein liquid chromatography. The specific linoleate oxygenase activity of the final enzyme preparation was 1300 nkat per mg protein at pH 6.8 and 25 degrees C in the absence of any detergent. The enzyme oxygenated linoleic acid and alpha-linolenic acid at comparable rates, whereas gamma-linolenic acid, arachidonic acid, 11,14-eicosadienoic acid and 11,14,17-eicosatrienoic acid were poor substrates. Linoleic acid was converted to 9(S)-hydroperoxy-10E,12Z-octadecadienoic acid, whereas 5(S)-HpETE, 11(S)-HpETE and 8(S)-HpETE were identified as major oxygenation products from arachidonic acid. The tomato lipoxygenase did not react with either dilinoleyl phosphatidylcholine or the lipid extract from beef heart mitochondria. The possible biological importance of the reaction of tomato lipoxygenase with arachidonic acid is discussed.
- Published
- 1994
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