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1.
In rat liver mitochondria, the macrocyclic polyether, dibenzo-18-crown-6 (polyether XXVIII) inhibits the oxidation of NAD-dependent substrates, as stimulated by ADP, uncouplers and valinomycin plus K+. It does not inhibit the oxidation of succinate. It is concluded that polyether XXVIII inhibits electron transfer in the NADH-CoQ span of the respiratory chain. This is a process that is reversed by menadione. Inhibition of oxidation of NAD-dependent substrates in K+-depleted mitochondria induced by the polyether is reversed by concentrations of K+ higher than 60 mM, and also by Li+, a cation that does not complex with polyether XXVIII. As assayed by swelling mitochondria, reversal of the inhibition of electron transfer is accompanied by influx of monovalent cations. Polyether XXVIII also inhibits in submitochondrial particles the aerobic oxidation of NADH, but not that of succinate; this inhibition is also reversed by K+ at high concentrations, and Li+. The data are consistent with the hypothesis that a monovalent cation is required for maximal rates of electron transport in the NADH-CoQ span of the respiratory chain. 相似文献
2.
There are many data both in favor and against the use of lucigenin as a probe for superoxide anion (SA) in mitochondria, cells, and simple enzymatic systems. In the present work high concentrations (50-400 M) of lucigenin were used for continuous recording of rapid and reversible changes in the SA level in intact isolated mitochondria. The SA level in the presence of lucigenin rapidly and reversibly changed during the transition of the mitochondria from one functional state to another: under conditions of ATP synthesis from ADP and Pi, of Ca2+ accumulation, and of reverse electron transfer. Induction of a Ca2+,cyclosporin A-sensitive pore in mitochondria completely suppressed the lucigenin-derived chemiluminescence (LDC). The electron transfer in the Q-cycle of the respiratory chain complex III and high electric potential difference across the inner membrane of mitochondria were obligatory conditions for generation of a SA-dependent chemiluminescent signal. Based on our own and literature data, a scheme of LDC generation is suggested. The origin of superoxide anion detected in intact mitochondria with lucigenin is discussed. 相似文献
3.
The spectrum of bixin shifts to longer wavelengths upon addition to mitochondria and the subsequent addition of the substrate of the electron transport system causes the lowering of the absorption band. The substrate-dependent absorbance decrease is suppressed by both inhibitors of respiration and uncoupling reagents, but not by oligomycin. The fact that bixin acts as an inhibitor of the ATP-forming process associated with respiration of mitochondria is supported by the experimental results obtained by measuring oxygen consumption, incorporation of 32Pi into ATP, ejection of H+ into the medium, adenosine triphosphatase activity, and the rate of the reversal electron flow. The lowering of the absorption band of bixin plus mitochondria caused by substrate reflects the energized sate of mitochondria. 相似文献
4.
Peter Schönfeld Mariusz R. Więckowski Magdalena Lebiedzińska Lech Wojtczak 《BBA》2010,1797(6-7):929-938
Reverse electron transfer (RET) from succinate to NAD+ is known to be accompanied by high generation of reactive oxygen species (ROS). In contrast, oxidation of fatty acids by mitochondria, despite being a powerful source of FADH2, does not lead to RET-associated high ROS generation. Here we show that oxidation of carnitine esters of medium- and long-chain fatty acids by rat heart mitochondria is accompanied by neither high level of NADH/NAD+ nor intramitochondrial reduction of acetoacetate to β-hydroxybutyrate, comparable to those accompanying succinate oxidation, although it produces the same or higher energization of mitochondria as evidenced by high transmembrane potential. Also in contrast to the oxidation of succinate, where conversion of the pH difference between the mitochondrial matrix and the medium into the transmembrane electric potential by addition of nigericin results in a decrease of ROS generation, the same treatment during oxidation of octanoylcarnitine produces a large increase of ROS. Analysis of respiratory chain complexes by Blue Native polyacrylamide gel electrophoresis revealed bands that could tentatively point to supercomplex formation between complexes II and I and complexes II and III. However, no such association could be found between complex I and the electron transferring flavoprotein that participates in fatty acid oxidation. It is speculated that structural association between respective respiratory chain components may facilitate effective reverse electron transfer. 相似文献
5.
Omar Ortiz-Avila Carlos Alberto Sámano-García Elizabeth Calderón-Cortés Ismael H. Pérez-Hernández Ricardo Mejía-Zepeda Alain R. Rodríguez-Orozco Alfredo Saavedra-Molina Christian Cortés-Rojo 《Journal of bioenergetics and biomembranes》2013,45(3):271-287
Impaired complex III activity and reactive oxygen species (ROS) generation in mitochondria have been identified as key events leading to renal damage during diabetes. Due to its high content of oleic acid and antioxidants, we aimed to test whether avocado oil may attenuate the alterations in electron transfer at complex III induced by diabetes by a mechanism related with increased resistance to lipid peroxidation. 90 days of avocado oil administration prevented the impairment in succinate-cytochrome c oxidoreductase activity caused by streptozotocin-induced diabetes in kidney mitochondria. This was associated with a protection against decreased electron transfer through high potential chain in complex III related to cytochromes c?+?c 1 loss. During Fe2+-induced oxidative stress, avocado oil improved the activities of complexes II and III and enhanced the protection conferred by a lipophilic antioxidant against damage by Fe2+. Avocado oil also decreased ROS generation in Fe2+-damaged mitochondria. Alterations in the ratio of C20:4/C18:2 fatty acids were observed in mitochondria from diabetic animals that not were corrected by avocado oil treatment, which yielded lower peroxidizability indexes only in diabetic mitochondria although avocado oil caused an augment in the total content of monounsaturated fatty acids. Moreover, a protective effect of avocado oil against lipid peroxidation was observed consistently only in control mitochondria. Since the beneficial effects of avocado oil in diabetic mitochondria were not related to increased resistance to lipid peroxidation, these effects were discussed in terms of the antioxidant activity of both C18:1 and the carotenoids reported to be contained in avocado oil. 相似文献
6.
A new theory for the electron transfer by the non-adiabatic process is formulated taking into account the origin shift and the frequency change of the vibration. The resultant formulas are quite similar to those of Jortner (Jortner, J. (1976) J. Chem. Phys. 64, 4860–4867) except that the free energy gap ΔG is used instead of the energy gap ΔE. By applying this theory to the photosynthetic electron transfer, the role of the remarkable temperature dependence of the electron transfer from cytochrome to P+ in Chromatium vinosum and the experimental data were reproduced very well using a small value of the coupling strength in contrast with the previous theory. This implies that proteins play a role to exclude many of the solvent molecules from the region of the electron transfer reaction between the donor and acceptor molecules. The negative activation process in the back electron transfer from Q?A to P+, the very slow back electron transfer from I? to P+ and the solvent isotope effect on the cytochrome oxidation are also successfully explained by this new theory. It is shown that even a qualitative conclusion as to the molecular parameters obtained from the temperature dependence of the electron transfer is different between the present theory and that of Jortner. 相似文献
7.
Dependency of the ATPase and 32 P--ATP exchange reaction of mitochondria on K + and electron transport 总被引:1,自引:0,他引:1
A Gómez-Puyou F Sandoval E Chávez D Freites M T De Gómez-Puyou 《Archives of biochemistry and biophysics》1972,153(1):215-225
The 2,4-dinitrophenol-stimulated ATPase activity and the 32P-ATP exchange reaction has been studied in rat liver mitochondria having less than 15 nmoles of K+ per milligram of protein. With 200 mm sucrose in the incubation media, the permeation of K+ and an oxidizable substrate is required for maximal stimulation of ATPase activity by 2,4-dinitrophenol. In these conditions, the 2,4-dinitrophenol-stimulated ATPase is inhibited by antimycin, acetate and mersalyl and depends to a certain extent on the rate of electron transport. The 32P-ATP exchange reaction of mitochondria with a low content of K+ also requires K+ permeation and is inhibited by antimycin, cyanide, 2,4-dinitrophenol, and acetate. The results suggest that the entrance of ATP into the mitochondria is compulsory linked to K+ uptake in a process that depends on a negative internal potential. 相似文献
8.
Mitochondria from skeletal muscle, heart and liver of strain 129/ReJ-dy dystrophic mice and their littermate controls were characterized with respect to their respiratory and phosphorylating activities. Skeletal muscle mitochondria from dystrophic mice showed significantly lower state 3 respiratory rates than controls with both pyruvate + malate and succinate as substrates (P < 0.01). ADP/O and Ca2+/O ratios were found to be normal. A decreased rate of NADH oxidation (0.01 <P < 0.05) by sonicated mitochondrial suspensions from dystrophic mice was also seen. High respiratory rates with ascorbate + phenazine methosulfate as substrates indicated that cytochrome oxidase was not rate limiting in the oxidation of either pyruvate + malate or succinate. Skeletal muscle mitochondria from dystrophic mice showed no deficiency in any of the cytochromes or coenzyme Q. Mg2+-stimulated ATPase activity was higher in dystrophic muscle mitochondria than in controls, but basal and oligomycin-insensitive activities were virtually identical to those of controls. A significant reduction in the intramitochondrial NAD+ content (0.01 <P < 0.02) was seen in dystrophic skeletal muscle as compared to controls. Heart mitochondria from dystrophic mice showed similar, though less extensive abnormalities while liver mitochondria were essentially normal. We concluded from these results that skeletal muscle mitochondria from strain 129 dystrophic mice possess impairments in substrate utilization which may result from (1) an abnormality in the transfer of electrons on the substrate side of coenzyme Q in the case of succinate oxidation; (2) a defect on the path of electron flow from NADH to cytochrome c, and (3) a deficiency of NAD+ in the case of NAD+-linked substrates. 相似文献
9.
Nohl H Gille L Kozlov A Staniek K 《Redox report : communications in free radical research》2003,8(3):135-141
The bioenergetic properties of mitochondria in combination with the high turnover rate of dioxygen qualify these organelles for the formation of reactive oxygen species (ROS). The assumption that mitochondria are the major intracellular source of ROS was essentially based on in vitro experiments with isolated mitochondria. The transfer of these data to the living cell may, however, be incorrect. Artefacts due to the preparation procedure or inadequate detection methods of ROS may lead to false positive results. Inhomogeneous results were found to be due to an interaction of the detection system with components of the respiratory chain which could be avoided by a recently developed non-invasive method. One of the most critical electron transfer steps in the respiratory chain is the electron bifurcation from ubiquinol to the cytochrome bc(1) complex. This electron bifurcation requires the free mobility of the head domain of the Rieske iron-sulfur protein. Inhibition of electron bifurcation by antimycin A causes leakage of single electrons to oxygen which results in the release of ROS. Hindrance of electron bifurcation was also observed following alterations of the physical state of membrane phospholipids in which the cytochrome bc(1) complex is inserted. Irrespective of whether the fluidity of the membrane was elevated or decreased, electron flow rates to the Rieske iron-sulfur protein were drastically reduced. Concomitantly superoxide radicals were released from these mitochondria, strongly suggesting the involvement of the ubiquinol/cytochrome bc(1) redox couple in this process. 相似文献
10.
Membrane energization at subzero temperatures: calcium uptake and oxonol-V responses 总被引:1,自引:0,他引:1
The use of aprotic solvents for preserving the electron transport properties of mitochondria at subzero temperatures is based upon the use of binary water and ethylene glycol mixtures or upon ternary and quaternary mixtures that include dimethyl sulfoxide and the lower aliphatic alcohols. In order to better preserve the respiratory control properties of mitochondria at subzero temperatures, detailed studies have been made of the effects of these mixtures on the respiratory control and electron transport from NADH or succinate of mitochondrial preparations. It is found that ADP is not metabolized at a measurable rate below 0 °C, but that Ca2+ is rapidly taken up and can thus be used to assay respiratory control ratios down to ?8 °C. In the region below ?8 °C the charge-sensitive probe oxonol-V has been used to evaluate energy coupling. By using Ca2+ to stimulate respiration at 0 °C good results are obtained with ethylene glycol/water alone and optimal results are obtained with a quaternary mixture. A mixture that freezes at ?21 °C gives about 50% inhibition of the respiratory control ratio for electron transport at 0 °C with NADH or succinate as substrates. The mixtures permit low-temperature studies of mitochondrial functions under conditions of minimal respiratory rate, including the kinetics of electron transfer reactions, the formation of intermediate compounds, and the rapid freeze-trapping of mitochondrial reactions for analytical chemistry or 31P NMR. 相似文献
11.
Ralph E. Bottenus Priscilla I. Spach Stephen Filus Carol C. Cunningham 《Biochemical and biophysical research communications》1982,105(4):1368-1373
Male rats were administered an ethanol-containing diet for 31 days during which time they demonstrated fatty liver. Mitochondria and submitochondrial particles were prepared from their livers (ethanol mitochondria, ethanol submitochondrial particles) and from their pair-fed partners (control mitochondria, control submitochondrial particles). The H+/coupling site ratio was not significantly different in ethanol and control mitochondria with succinate as electron donor. A 13% decrease in the H+/coupling site ratio was observed in ethanol mitochondria, however, when β-hydroxybutyrate was used as substrate. The rate of ATP-Pi exchange was decreased significantly in both ethanol mitochondria and submitochondrial particles as compared to control preparations. These observations demonstrate ethanol-elicited decreases in energy conservation in the site I region of the electron transport chain and in the activity of the ATP synthetase complex. 相似文献
12.
Inorganic phosphate stimulates the release of Mg++ from liver mitochondria, depending on concentration; a concentration as low as 0.1 mM phosphate is already effective. The process is dependent on the electron transfer of the respiratory chain, and its rate is highest under conditions of endogenous respiration and with ascorbate and TMPD as substrates, respectively. The phosphate stimulated release of Mg++ is followed, with a pronounced delay, by a Ca++ efflux and a swelling of mitochondria. Addition of EGTA strongly reduced the rate of Mg++ liberation in the presence and absence of inorganic phosphate. Exogenous Ca++ is able to abolish the EGTA effect. ADP and ATP inhibit the phosphate stimulated release of Mg++. Phosphoenol pyruvate and free fatty acids enhance the rate of Mg++ and Ca++ efflux from the mitochondria. The results permit the conclusion that inorganic phosphate, Ca++ and various metabolites of the cell metabolism influence the Mg++ distribution between the extra- and intramitochondrial space, thus controlling the permeability of the mitochondrial inner membrane for monovalent cations. 相似文献
13.
《Plant Science Letters》1978,11(3-4):345-350
Plant mitochondria oxidize exogenous NADH via an external NADH dehydrogenase; little is known about the regulation of this dehydrogenase. Data presented here show that citrate caused a non-competitive inhibition of NADH oxidation. This inhibition can be reversed by Ca2+. It is postulated that citrate causes the inhibition by extracting Ca2+ from the NADH-ubiquinone reductase. Citrate caused a more powerful effect if added before rather than after the NADH. This suggests that the Ca2+ is less easily removed in the presence of NADH due to a conformational change in the membrane. Experimentals using a series of electron acceptors suggest that citrate prevents electron transfer between the flavoprotein and the ubiquinone. It is suggested that calcium facilitates electron flux between these components by binding the flavoprotein to the membrane. 相似文献
14.
《Autophagy》2013,9(3):405-408
Mitochondria are the primary site of energy production in animal cells. In mitochondria, the flow of electrons through the electron transport chain creates a potential difference across the inner membrane, which is utilized for ATP production. However, due to inherent inefficiencies in electron transport, reactive oxygen species are also produced, which damage mitochondrial proteins and nucleic acids, and impair mitochondrial function.1 Decreased mitochondrial function causes increased reactive oxygen species generation, a decline in cellular function, and potentially cell death.2 Therefore, to maintain cellular homeostasis, mechanisms have evolved to selectively eliminate defective mitochondria.3 Mitochondria are constantly undergoing cycles of fission and fusion, and this process appears to have a role in mitochondrial quality control. Following fission, daughter mitochondria are produced, which can differ in their membrane polarization. Depolarized mitochondria are less likely to undergo subsequent fusion, and more likely to undergo autophagic clearance.4 As would be predicted, given the potential for cytochrome c release, depolarization is a powerful stimulus for mitochondrial clearance. Depolarization causes recruitment of the E3 ubiquitin ligase Parkin to mitochondria, which is required for their subsequent engulfment by autophagosomes.5 Macroautophagy pathways also appear to have a role, as hepatocytes deficient for the E1-like enzyme Atg7 accumulate abnormal mitochondria.6 Finally, recent studies in a developmental model have yielded insight into this process. Newly-formed erythrocytes, also known as reticulocytes, eliminate their entire cohort of mitochondria during development.7 This process depends on the mitochondrial protein NIX, is partially dependent on autophagy, and is independent of mitochondrial depolarization.8-10 Here we describe the use of reticulocytes to study mitochondrial clearance. 相似文献
15.
I. MAX MØLLER 《Physiologia plantarum》1978,42(2):157-162
Mitochondria were isolated from 7-day-old wheat roots (Triticum vulgare Vill. cv. Svenno Spring Wheat) grown in either a full-strength culture medium (100%) or in the same medium diluted 100 times (1%). Outer membrane integrity was assayed using the cytochrome c reduction assay. This indicated about 20% damage. Using an oxygen electrode the respiration of the mitochondria was measured with either malate or succinate as the substrate (both 40 mM). KCN (3 mM) and salicylhydroxamic acid (SHAM, 1 mM) were used as inhibitors. The properties of the isolated mitochondria (STATE 3 rate, ADP/O ratio, and KCN-sensitivity) depend upon the ionic concentration of the growth medium of the roots. In the mitochondria isolated from roots grown in the 1% medium (1% mitochondria) there is a synergistic effect of KCN and SHAM. This means that electrons can be shifted from one pathway to the other when only one of the inhibitors is added. This flexibility between the electron pathways is almost nil in the mitochondria isolated from roots grown in the 100% medium (100% mitochondria). The maximal capacity of the alternative electron pathway (= rate in the presence of KCN) is higher in 1% (40 nmol O2 min?1 (mg protein)?1) than in 100% mitochondria (20 nmol O2 min?1 (mg protein)?1. In 100% mitochondria the alternative pathway seems to be operating at maximal capacity in the absence of KCN with both substrates and in both STATES 3 and 4. In 1% mitochondria the alternative pathway functions at >50% of its capacity in the absence of KCN. 相似文献
16.
The mechanisms of hemoglobin autoxidation. Evidence for proton-assisted nucleophilic displacement of superoxide by anions 总被引:2,自引:0,他引:2
W J Wallace J C Maxwell W S Caughey 《Biochemical and biophysical research communications》1974,57(4):1104-1110
Human oxyhemoglobin (HbO2) in the presence of excess nucleophile (e.g., N3?, SCN?, F?, Cl?) is shown by visible and Soret spectra to form cleanly the oxidized metHb with the nucleophile as ligand. The rates, sensitive to pH and to both the concentration and the nucleophilicity of anionic nucleophile (N?), follow the rate law: rate = k[HbO2][N?][H+]. This autoxidation process thus appears to involve the nucleophilic displacement of superoxide from a protonated intermediate and can reasonably account for normal metHb formation in the erythrocyte where chloride can serve as the nucleophile. MetHb formation due to electron transfer agents (e.g. nitrite) which are normally not present can follow a different course such as direct electron transfer to bound dioxygen to form iron (III) peroxide. Abnormal amino acids or denaturation can provide increased access of nucleophile or electron transfer reactant and thus promote autoxidation. 相似文献
17.
Mauricio CortesJessica T Carney Jonathon D OppenheimerKaren E Downey Scott D Cummings 《Inorganica chimica acta》2002,333(1):148-151
The luminescent complex [Pt(terpy)OH]BF4 undergoes photoinduced electron transfer reactions with phenyl amine electron donors and nitrophenyl electron acceptors. Stern-Volmer analysis of the quenching of metal-to-ligand charge transfer phosphorescence (3MLCT) was used to calculate bimolecular rate constants for electron transfer. Rate constants vary from 108 to >1010 M−1 s−1, depending on the thermodynamic driving force of the electron transfer reaction, with rate constants indicating that [Pt(terpy)OH]BF4* is a powerful photo-oxidant. Aromatic triplet energy acceptors can also quench the 3MLCT emission. 相似文献
18.
I. V. Zhigacheva E. B. Burlakova A. G. Shugaev I. P. Generozova S. G. Fattakhov A. I. Konovalov 《Biochemistry (Moscow) Supplemental Series A: Membrane and Cell Biology》2008,2(3):231-236
The addition of the organophosphorous plant growth regulator Melaphen (4 × 10?12 M) to the incubation medium increases the maximum rate of oxidation of NAD-dependent substrates in rat liver and sugar beet root mitochondria. In addition, Melaphen stimulates electron transport during oxidation of succinate by rat liver mitochondria, but has no effect on the rate of this substrate oxidation in sugar beet root mitochondria. In storage organs of plants, the rate of oxidation of NAD-dependent substrates by mitochondria is relatively low. By stimulating the activity of NAD-dependent dehydrogenases, Melaphen stimulates energy metabolism in the cells and manifests adaptogenic activity by accelerating the germination of seeds. Melaphen does not influence the fluorescence of lipid peroxidation (LPO) products in mitochondria non-exposed to stress, but decreases 1.5–2 fold the LPO fluorescence in rat liver mitochondria exposed to cold stress and artificially “aged” sugar beet root mitochondria. Besides, Melaphen increases the rate of electron transport in a terminal site of respiratory chains of plant and animal mitochondria and decreases LPO. The data obtained testify to antistress activity of Melaphen. 相似文献
19.
Active oxidative decarboxylation of malate by mitochondria isolated from L-1210 ascites tumor cells 总被引:3,自引:0,他引:3
Mitochondria from L-1210 mouse ascites tumor show a very high rate of oxidation of L-malate in comparison with mitochondria from normal tissues. They were found to contain large amounts of malic enzyme (E.C.1.1.1.39) catalyzing oxidative decarboxylation of L-malate to pyruvate. Malic enzyme in extracts of tumor mitochondria requires Mn2+ or Mg2+, utilizes either NAD+ or NADP+ as electron acceptor, and shows positive cooperativity in binding of L-malate. These observations suggest that L-1210 tumor mitochondria actively convert excess tricarboxylate cycle intermediates and their precursors into pyruvate for further oxidation. 相似文献
20.
Ernesto Carafoli Marina Gavilanes Hubert Affolter Marietta Tuena de Gómez-Puyou Armando Gómez-Puyou 《Cell calcium》1980,1(4):255-265
The ATP-supported Ca2+ uptake of heart and liver mitochondria preincubated in conditions in which electron transport had either been prevented by rotenone or antimycin, or induced by oxidizable substrates, has been studied. Mitochondria preincubated with respiratory inhibitors accumulate Ca2+ less efficiently than mitochondria preincubated with oxidizable substrates. The difference correlates with the degree of activation of the oligomycin-sensitive ATPase. The results indicate that the rate at which mitochondria take up Ca2+ in the ATP-supported system may be controlled by the reversible asociation of the inhibiting peptide (Pullman,. and Monroy, J. Biol. Chem., 238, 3762–3769) with the ATPase complex. Since this process appears to be modulated by the transmembrane electrochemical gradient, the latter may regulate the uptake of Ca2+ in a hitherto undescribed way. 相似文献