scholarly journals Low-level chemiluminescence of bovine heart submitochondrial particles

1980 ◽  
Vol 186 (3) ◽  
pp. 659-667 ◽  
Author(s):  
Enrique Cadenas ◽  
Alberto Boveris ◽  
Britton Chance
Keyword(s):  
Lipid Peroxidation ◽  
Cytochrome C ◽  
Light Emission ◽  
Cumene Hydroperoxide ◽  
Radical Reactions ◽  
Submitochondrial Particles ◽  
Butyl Hydroperoxide ◽  
Low Level ◽  
Bovine Heart ◽  
Β Carotene

Submitochondrial particles from bovine heart mitochondria showed low-level chemiluminescence when supplemented with organic hydroperoxides. Chemiluminescence seems to measure integratively radical reactions involved in lipid peroxidation and related processes. Maximal light-emission was about 1500 counts/s and was reached 2–10min after addition of hydroperoxides. Ethyl hydroperoxide, cumene hydroperoxide and t-butyl hydroperoxide were effective in that order. Antimycin and rotenone increased chemiluminescence by 50–60%; addition of substrates, NADH and succinate did not produce marked changes in the observed chemiluminescence. Cyanide inhibited chemiluminescence; half-maximal inhibitory effect was obtained with 0.03mm-cyanide and the inhibition was competitive with respect to t-butyl hydroperoxide. Externally added cytochrome c (10–20μm) had a marked stimulatory effect on chemiluminescence, namely a 12-fold increase in light-emission of antimycin-inhibited submitochondrial particles. Stimulation of hydroperoxide-induced chemiluminescence of submitochondrial particles by cytochrome c was matched by a burst of O2 consumption. O2 is believed to participate in the chain radical reactions that lead to lipid peroxidation. Superoxide anion seems to be involved in the chemiluminescence reactions as long as light-emission was 50–60% inhibitible by superoxide dismutase. Singlet-oxygen quenchers, e.g. β-carotene and 1,4-diazabicyclo[2,2,2]-octane, affected light-emission. β-Carotene was effective either when incorporated into the membranes or added to the cuvette. The present paper suggests that singlet molecular oxygen is mainly responsible for the light-emission in the hydroperoxide-supplemented submitochondrial particles.

scholarly journals Low-level chemiluminescence of hydroperoxide-supplemented cytochrome c

1980 ◽  
Vol 187 (1) ◽  
pp. 131-140 ◽  
Author(s):  
Enrique Cadenas ◽  
Alberto Boveris ◽  
Britton Chance
Keyword(s):  
Cytochrome C ◽  
Singlet Oxygen ◽  
Light Emission ◽  
Cumene Hydroperoxide ◽  
Present System ◽  
Butyl Hydroperoxide ◽  
Low Level ◽  
Radical Trap ◽  
Organic Hydroperoxides ◽  
Ferricytochrome C

Ferricytochrome c showed low-level chemiluminescence, with a light-emission measured of about 1×103–3×103 counts/s, when supplemented with organic hydroperoxides. Tertiary hydroperoxides (cumene hydroperoxide and t-butyl hydroperoxide) showed a saturation behaviour at about 5mm-hydroperoxide, whereas primary hydroperoxides showed a quadratic dependence on the hydroperoxide concentration. Chemiluminescence depended linearly on cytochrome c concentration, and optimal light-emission was observed at [t-butyl hydroperoxide]/[ferricytochrome c] ratios of 160–500. Hydroperoxide-supplemented ferricytochrome c consumed O2 at a rate of 1.0μmol/min per μmol of cytochrome c; the rate of O2 uptake was linearly related to the concentration of cytochrome c. The Soret absorption band of ferricytochrome c decreased about 64% after incubation with t-butyl hydroperoxide, whereas the 530nm band was almost totally abolished. Light-emission was (a) inhibited competitively by cyanide. (b) inhibited by singlet-oxygen quenchers (e.g. β-carotene), scavengers (e.g. dimethylfuran) and traps (e.g. histidine and tryptophan) and (c) increased by singlet-oxygen-chemiluminescence enhancer 1,4-diazabicyclo[2.2.2]-octane. Superoxide dismutase had no effect on the present system. The participation of free radicals is suggested by the effect of the radical trap 2,5-di-t-butylquinol. Singlet-oxygen dimol emission seems to be mainly responsible for the observed light-emission; a mechanism that can account for the major part of the present experimental observations is proposed.

scholarly journals Alteration of inner-membrane components and damage to electron-transfer activities of bovine heart submitochondrial particles induced by NADPH-dependent lipid peroxidation

1982 ◽  
Vol 202 (1) ◽  
pp. 97-105 ◽  
Author(s):  
H Narabayashi ◽  
K Takeshige ◽  
S Minakami
Keyword(s):  
Lipid Peroxidation ◽  
Electron Transfer ◽  
Cytochrome C ◽  
Succinate Dehydrogenase ◽  
Sodium Dodecyl ◽  
Submitochondrial Particles ◽  
Inner Membrane ◽  
Cytochrome C Reductase ◽  
Bovine Heart ◽  
Membrane Components

We investigated the changes of the inner-membrane components and the electron-transfer activities of bovine heart submitochondrial particles induced by the lipid peroxidation supported by NADPH in the presence of ADP-Fe3+. Most of the polyunsaturated fatty acids were lost as a result of the peroxidation, and phospholipids were changed to polar species. Ubiquinone was also modified to polar substances as the peroxidation proceeded. Sodium dodecyl sulphate/polyacrylamide-gel electrophoresis showed the disappearance of 27000-Mr and 30000-Mr proteins and the appearance of highly polymerized substances. Flavins and cytochromes were not diminished, but the respiratory activity was lost. The reactions of NADH oxidase and NADH-cytochrome c reductase were most sensitive to the peroxidation, followed by those of succinate oxidase and succinate-cytochrome c reductase. Succinate dehydrogenase and duroquinol-cytochrome c reductase were inactivated by more extensive peroxidation, but cytochrome c oxidase was only partially inactivated. NADH-ferricyanide reductase was not inactivated. The pattern of the inactivation indicated that the lipid peroxidation affected the electron transport intensively between NADH dehydrogenase and ubiquinone, and moderately at the succinate dehydrogenase step and between ubiquinone and cytochrome c.

scholarly journals Inhibition and inactivation of NADH-cytochrome c reductase activity of bovine heart submitochondrial particles by the iron(III)-adriamycin complex

1990 ◽  
Vol 265 (3) ◽  
pp. 865-870 ◽  
Author(s):  
B B Hasinoff
Keyword(s):  
Cytochrome C ◽  
Reductase Activity ◽  
Inner Mitochondrial Membrane ◽  
Submitochondrial Particles ◽  
Cytochrome C Reductase ◽  
Fast Phase ◽  
Bovine Heart ◽  
Direct Binding ◽  
Nadh Cytochrome

The NADH-cytochrome c reductase activity of bovine heart submitochondrial particles was found to be slowly (half-time of 16 min) and progressively lost upon incubation with the Fe2(+)-adriamycin complex. In addition to this slow progressive inactivation seen on incubation, a reversible fast phase of inhibition was also seen. However, if EDTA was added to the incubation mixture within 15 s, the slow progressive loss in activity was largely preventable. Separate experiments indicated that EDTA removed about one-half of the iron from the Fe2(+)-adriamycin complex in about 40 s. These results indicated the requirement for iron for the inactivation process. Since the Vmax. for the fast phase of inhibition was decreased by the inhibitor, the inhibition pattern was similar to that seen for uncompetitive or mixed-type inhibition. The direct binding of both Fe3(+)-adriamycin and adriamycin to submitochondrial particles was also demonstrated, with the Fe3(+)-adriamycin complex binding 8 times more strongly than adriamycin. Thus binding of Fe3(+)-adriamycin to the enzyme or to the inner mitochondrial membrane with subsequent generation of oxy radicals in situ is a possible mechanism for the Fe3(+)-adriamycin-induced inactivation of respiratory enzyme activity.

Lipid peroxidation by bovine heart submitochondrial particles stimulated by 1,1′-dimethyl-4,4′-bipyridylium dichloride (paraquat)

1983 ◽  
Vol 32 (1) ◽  
pp. 13-19 ◽  
Author(s):  
Takeyoshi Sata ◽  
Koichiro Takeshige ◽  
Ryoichi Takayanagi ◽  
Shigeki Minakami
Keyword(s):  
Lipid Peroxidation ◽  
Submitochondrial Particles ◽  
Bovine Heart

scholarly journals NADH- and NADPH-dependent lipid peroxidation in bovine heart submitochondrial particles. Dependence on the rate of electron flow in the respiratory chain and an antioxidant role of ubiquinol

1980 ◽  
Vol 192 (3) ◽  
pp. 853-860 ◽  
Author(s):  
R Takayanagi ◽  
K Takeshige ◽  
S Minakami
Keyword(s):  
Lipid Peroxidation ◽  
Respiratory Chain ◽  
Electron Flow ◽  
Antimycin A ◽  
Submitochondrial Particles ◽  
Bovine Heart ◽  
Malondialdehyde Formation ◽  
Low Rate ◽  
Liquid Peroxidation

Malondialdehyde formations by bovine heart submitochondrial particles supported by NADH or NADPH in the presence of ADP and FeCl3 was studied. The NADH-dependent reaction was maximal at very low rate of electron input from NADH to the respiratory chain and it decreased when the rate became high. The reaction was stimulated by rotenone and inhibited by antimycin A when the input was fast, whereas it was not affected by the inhibitors when the input was slow. The input rate of the electrons from NADPH was also so low that the reaction supported by NADPH was not affected by the inhibitors. Most of the endogenous ubiquinone in the particles treated with antimycin A was reduced by NADH even in the presence of ADP-Fe3+ chelate, but uniquinone was not reduced by NADPH when ADP-Fe3+ was present. Succinate strongly inhibited both NADH- and NADPH-dependent lipid peroxidation. The inhibition was abolished when uniquinone was removed from the particles, and it appeared again when uniquinone was reincorporated into the particles. Reduced uniquinone-2 also inhibited the peroxidation, but duroquinol, which reduces cytochrome b without reducing endogenous uniquinone, did not. Thus the malondialdehyde formation appeared to be inversely related to the extent of the reduction of endogenous uniquinone. These observations suggest that both NADH- and NADPH-dependent liquid-peroxidation reactions are closely related to the respiratory chain and that the peroxidation is controlled by the concentration of reduced ubiquinone.

scholarly journals Kinetics of inhibition of purified and mitochondrial cytochrome c oxidase by psychosine (β-galactosylsphingosine)

1993 ◽  
Vol 290 (1) ◽  
pp. 139-144 ◽  
Author(s):  
C E Cooper ◽  
M Markus ◽  
S P Seetulsingh ◽  
J M Wrigglesworth
Keyword(s):  
Cytochrome C ◽  
Cytochrome Oxidase ◽  
Cytochrome C Oxidase ◽  
Rat Liver ◽  
Liver Mitochondria ◽  
Slow Phase ◽  
Rat Liver Mitochondria ◽  
Mitochondrial Cytochrome ◽  
Submitochondrial Particles ◽  
Bovine Heart

1. Psychosine (beta-galactosylsphingosine) is the toxic agent in Krabbe's disease (globoid cells leukodystrophy). It inhibits purified bovine heart mitochondrial cytochrome c oxidase; there is a rapid phase of inhibition (complete within 10-15 s) and a slower phase (complete within 10-15 min). Both phases are also seen in rat liver mitochondria. IC50 is about 200 microM psychosine in the purified enzyme and less than 20 microM in mitochondria. Psychosine inhibition is due to binding to cytochrome oxidase, not cytochrome c. 2. Bovine heart submitochondrial particles show inhibition similar to rat liver mitochondria. However, although proteoliposomes containing bovine heart cytochrome oxidase show an identical fast phase, they have no noticeable slow phase of inhibition. Addition of phospholipid liposomes to submitochondrial particles relieved the majority of psychosine inhibition, consistent with the removal of those molecules binding in the slow phase. Psychosine can inhibit cytochrome oxidase molecules facing in either direction in proteoliposomes and submitochondrial particles, suggesting that it can rapidly interact with both sides of a membrane when added externally. 3. At high ionic strength, the presence of psychosine decreases the Vmax. of cytochrome oxidase with little effect on the Km for cytochrome c. This non-competitive inhibition suggests that the psychosine-enzyme complex is kinetically inactive and not labile over the time course of the assay. Psychosine does not inhibit the reduction of haem a or haem a3 by artificial electron donors, but does inhibit the reduction of haem a by cytochrome c.

Effect of β-Carotene and Canthaxanthin ontert-Butyl Hydroperoxide-Induced Lipid Peroxidation in Murine Normal and Tumor Thymocytes

1996 ◽  
Vol 325 (2) ◽  
pp. 145-151 ◽  
Author(s):  
Paola Palozza ◽  
Chiara Luberto ◽  
Paola Ricci ◽  
Elisabetta Sgarlata ◽  
Gabriella Calviello ◽  
...  
Keyword(s):  
Lipid Peroxidation ◽  
Butyl Hydroperoxide ◽  
Β Carotene
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