RESUMEN
Liver ischemia-reperfusion injury is still an open problem in many clinical circumstances, including surgery and transplantation. This study investigates how mitochondrial structure, mass and oxidative phosphorylation change and may be preserved during a brief period of ischemia followed by a long period of reperfusion, an experimental model that mimics the condition to which a liver is exposed during transplantation. Livers were explanted from rats and exposed for 24 h to three different oxygen availability conditions at 4 °C. Mitochondrial mass, respiration, oxidative phosphorylation (OXPHOS), and levels of OXPHOS complexes were all significantly altered in livers stored under the currently used preservation condition of normoxia. Remarkably, liver perfusion with hyperoxic solutions fully preserved mitochondrial morphology and function, suggesting that perfusion of the graft with hyperoxic solution should be considered in human transplantation.
Asunto(s)
Hiperoxia/metabolismo , Hígado/metabolismo , Mitocondrias Hepáticas/metabolismo , Fosforilación Oxidativa , Consumo de Oxígeno , Animales , Humanos , Hiperoxia/patología , Isquemia/metabolismo , Isquemia/patología , Hígado/patología , Trasplante de Hígado , Mitocondrias Hepáticas/patología , Ratas , Ratas Sprague-Dawley , ReperfusiónRESUMEN
The supra-molecular assembly of the main respiratory chain enzymatic complexes in the form of "super-complexes" has been proved by structural and functional experimental evidence. This evidence strongly contrasts the previously accepted Random Diffusion Model stating that the complexes are functionally connected by lateral diffusion of small redox molecules (i.e. Coenzyme Q and cytochrome c). This review critically examines the available evidence and provides an analysis of the functional consequences of the intermolecular association of the respiratory complexes pointing out the role of Coenzyme Q and of cytochrome c as channeled or as freely diffusing intermediates in the electron transfer activity of their partner enzymes.
Asunto(s)
Transporte de Electrón , Mitocondrias/metabolismo , Fosforilación Oxidativa , Consumo de Oxígeno , Animales , Citocromos c/química , Citocromos c/metabolismo , Cinética , Mitocondrias/enzimología , Modelos Moleculares , Complejos Multienzimáticos/química , Complejos Multienzimáticos/metabolismo , NADH NADPH Oxidorreductasas/química , NADH NADPH Oxidorreductasas/metabolismo , Oxidorreductasas/química , Oxidorreductasas/metabolismo , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Ubiquinona/química , Ubiquinona/metabolismoRESUMEN
Asbestos fibers, such as chrysotile and crocidolite, are known to have cytotoxic effects on different cell types. In vivo exposure to asbestos fibers can induce both fibrotic and malignant lung diseases , however, the mechanisms linking exposure to the subsequent development of the diseases are unknown. Numerous investigations suggest the involvement of reactive oxygen species (ROS). ROS are known to damage biological macromolecules including proteins, cell membrane lipids and nucleic acids; alterations of these essential cellular components can alter cell function and can drive the cell to neoplastic transformation or to cell death. Because the mitochondrial respiratory chain is an important source of ROS and RNS (reactive nitogen species) in the cells, we have investigated the effects of aqueous extracts of asbestos (natural and synthetic) fibers on some mitochondrial activities. Our data show that crocidolite fibers release substances in solution that may interfere directly with the mitochondrial cytochrome oxidase complex. Moreover, the calcium ions released from these fibers induce opening of the permeability transition pore of the inner membrane leading to a possible cytotoxic effect due to the release of apoptotic factors normally localized in the mitochondrial intermembrane space. In addition, crocidolite extracts enhance the mitochondrial production of ROS. No significant biochemical effects are exerted by chrysotile, either natural or synthetic, on isolated mitochondria. Nevertheless, all asbestos fibers tested induce morphological alterations visualized by transmission electron microscopy and morphometric analysis.
Asunto(s)
Asbesto Crocidolita/toxicidad , Mitocondrias/efectos de los fármacos , Animales , Asbesto Crocidolita/química , Calcio/metabolismo , Permeabilidad de la Membrana Celular/efectos de los fármacos , Complejo IV de Transporte de Electrones/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Mitochondria are deeply involved in the production of reactive oxygen species through one-electron carriers in the respiratory chain; mitochondrial structures are also very susceptible to oxidative stress as evidenced by massive information on lipid peroxidation, protein oxidation, and mitochondrial DNA (mtDNA) mutations. Oxidative stress can induce apoptotic death, and mitochondria have a central role in this and other types of apoptosis, since cytochrome c release in the cytoplasm and opening of the permeability transition pore are important events in the apoptotic cascade. The discovery that mtDNA mutations are at the basis of a number of human pathologies has profound implications: maternal inheritance of mtDNA is the basis of hereditary mitochondrial cytopathies; accumulation of somatic mutations of mtDNA with age has represented the basis of the mitochondrial theory of ageing, by which a vicious circle is established of mtDNA damage, altered oxidative phosphorylation and overproduction of reactive oxygen species. Experimental evidence of respiratory chain defects and of accumulation of multiple mtDNA deletions with ageing is in accordance with the mitochondrial theory, although some other experimental findings are not directly ascribable to its postulates.
Asunto(s)
Envejecimiento/metabolismo , Mitocondrias/metabolismo , Estrés Oxidativo , Animales , Muerte Celular , Daño del ADN , ADN Mitocondrial/genética , ADN Mitocondrial/metabolismo , Metabolismo Energético , Radicales Libres/metabolismo , Humanos , Mutación , Enfermedades Neurodegenerativas/metabolismo , Especies Reactivas de Oxígeno/metabolismoRESUMEN
This review considers the interaction of Complex I with different redox acceptors, mainly homologs and analogs of the physiological acceptor, hydrophobic Coenzyme Q. After examining the physical properties of the different quinones and their efficacy in restoring mitochondrial respiration, a survey ensues of the advantages and drawbacks of the quinones commonly used in Complex I activity determination and of their kinetic properties. The available evidence is then displayed on structure-activity relationships of various quinone compounds in terms of electron transfer activity and proton translocation, and the present knowledge is discussed in terms of the nature of multiple quinone-binding sites in the Complex.
Asunto(s)
NAD(P)H Deshidrogenasa (Quinona)/química , Quinonas/química , NAD(P)H Deshidrogenasa (Quinona)/metabolismo , Relación Estructura-ActividadRESUMEN
This investigation shows that the effects of general anesthetics previously observed in vitro on membrane fluidity and on enzymic activities and occurring at concentrations calculated to be clinically relevant can be reproduced in vivo in anesthetized animals. Anesthesia with 2-chlorophenyl-2-methylaminocyclohexanone (ketamine) induces a more fluid state of rat-brain synaptic and mitochondrial membranes, as shown by the rotational correlation times of the spin labels 16-doxylstearate and 5-doxylstearate. Changes in acetylcholinesterase activity, with a decrease in Vmax and no change in the Km for acetylcholine, closely follow the fluidity increase.
Asunto(s)
Acetilcolinesterasa/metabolismo , Encéfalo/efectos de los fármacos , Ketamina/farmacología , Fluidez de la Membrana/efectos de los fármacos , Animales , Encéfalo/enzimología , Óxidos N-Cíclicos/metabolismo , Cinética , Masculino , Membranas/efectos de los fármacos , Ratas , Ratas EndogámicasRESUMEN
A kinetic study on ubiquinol-cytochrome c reductase (EC 1.10.2.2) has been undertaken either in situ in KCN-inhibited mitochondria and submitochondrial particles, or in the isolated cytochrome b-c1 complex using ubiquinol-1 and exogenous cytochrome c as substrates. The steady-state two-substrate kinetics of the reductase appears to follow a general sequential mechanism, allowing calculation of a Km for ubiquinol-1 of 13.4 microM in mitochondria and of 24.6 microM in the isolated cytochrome b-c1 complex. At low concentrations of cytochrome c, however, the titrations as a function of quinol concentration appear biphasic both in mitochondria and in submitochondrial particles containing trapped cytochrome c inside the vesicle space, fitting two apparent Km values for ubiquinol-1. Relatively high antimycin-sensitive rates of ubiquinol-1-cytochrome c reductase have been found in submitochondrial particles: both the Vmax and the Km for ubiquinol-1 are, however, affected by the overall orientation of the particle preparation, i.e., by the reactivity of cytochrome c with its proper site. The turnover numbers corrected for particle orientation with respect to cytochrome c interaction are at least 2-fold higher in submitochondrial particles than in mitochondria. This is particularly evident using inside-out particles containing trapped cytochrome c in the vesicle space (and therefore reacting with its physiological site). A diffusion step for the quinol substrate appears to be rate limiting in mitochondria and can be removed by addition of deoxycholate, suggesting that the oxidation site of ubiquinol may be more exposed to the matrix side of the inner mitochondrial membrane.
Asunto(s)
Mitocondrias Cardíacas/enzimología , Mitocondrias/enzimología , Complejos Multienzimáticos/metabolismo , NADH NADPH Oxidorreductasas/metabolismo , Quinona Reductasas/metabolismo , Partículas Submitocóndricas/enzimología , Animales , Bovinos , Grupo Citocromo c/metabolismo , Ácido Desoxicólico/farmacología , Complejo III de Transporte de Electrones , CinéticaRESUMEN
We have devised a method to determine the true Km of membrane enzymes for hydrophobic substrates dissolved in lipid bilayers, and the lipid/water partition coefficients, by simple steady-state kinetic measurements at varying membrane phospholipid fractional volumes in the assay medium. The method has been applied to mitochondrial ubiquinol cytochrome c reductase, using short-chain ubiquinols as reductants at saturating cytochrome c. The partition coefficients of the quinols, as obtained by this method, are in good agreement with those determined directly by other procedures; Km values obtained by this method, when expressed as concentrations in the lipid bilayer, are in the millimolar range. The kinetics of the ubiquinol analog duroquinol are independent of phospholipid concentration, as expected from its partition coefficient close to unity.
Asunto(s)
Complejo III de Transporte de Electrones/metabolismo , Algoritmos , Animales , Bovinos , Cinética , Métodos , Mitocondrias Cardíacas/enzimología , Mitocondrias Cardíacas/ultraestructura , Partículas Submitocóndricas/enzimologíaRESUMEN
We have investigated in detail the effects of dibromothymoquinone (2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone, DBMIB) on the ubiquinol-cytochrome c reductase (cytochrome bc1 complex) from bovine heart mitochondria. The inhibitory action of DBMIB on the steady-state activity of the bc1 complex is related to the specific binding of the quinone to the purified enzymatic complex. At concentrations higher than 10 mol per mol of the enzyme, DBMIB is able to stimulate an antimycin-insensitive reduction of cytochrome c catalyzed by the bc1 complex. In accordance with kinetic data showing a competition by endogenous ubiquinone in the inhibitory action, DBMIB can be considered as a product-like inhibitor of the ubiquinol-cytochrome c reductase activity. The site of specific binding of dibromothymoquinone in the bc1 complex enables it to interact with the iron-sulphur center of the enzyme, as indicated by changes induced in the EPR spectrum of the center. However, the inhibitor also directly interacts with cytochrome b, promoting a fast chemical oxidation of the reduced heme center. In spite of these effects, DBMIB has been found not to exert significant effects on the first turnover of the fully oxidized bc1 complex, as monitored by the rapid reduction of both cytochromes b and c1 by ubiquinol-1. In the presence of antimycin, only a stimulation of cytochrome c1 reduction, in parallel to an enhanced cytochrome b reoxidation, is observed. Moreover, DBMIB does not affect the oxidant-induced extra cytochrome b reduction in the presence of antimycin. On the basis of the evidences suggesting a competition with the endogenous ubiquinone in the redox cycle of the bc1 complex, a model is proposed for the mechanism of DBMIB inhibition. Such model can also explain at the molecular level the redox bypass induced by dibromothymoquinone in the whole respiratory chain (Degli Esposti, M., Rugolo, M. and Lenaz, G. (1983) FEBS Lett. 156, 15-19).
Asunto(s)
Dibromotimoquinona/farmacología , Mitocondrias Cardíacas/enzimología , Complejos Multienzimáticos/antagonistas & inhibidores , NADH NADPH Oxidorreductasas/antagonistas & inhibidores , Quinona Reductasas/antagonistas & inhibidores , Quinonas/farmacología , Animales , Unión Competitiva , Bovinos , Grupo Citocromo b/metabolismo , Grupo Citocromo c/metabolismo , Citocromos c1/metabolismo , Dibromotimoquinona/metabolismo , Espectroscopía de Resonancia por Spin del Electrón , Complejo III de Transporte de Electrones , Cinética , Complejos Multienzimáticos/metabolismo , Oxidación-Reducción , Quinona Reductasas/metabolismo , Ubiquinona/farmacologíaRESUMEN
The circular dichroic spectrum of the mitochondrial cytochrome bc1 complex isolated from bovine heart has been resolved into the contributions from the prosthetic groups: cytochrome c1, the 'Rieske' iron-sulphur centre and the two b cytochromes. It is apparent that firstly, the circular dichroism (CD) properties of cytochrome c1 within the bc1 complex differ from those found in the isolated cytochrome c1 and secondly, both the oxidized and reduced b cytochromes exhibit an intense spectrum of bilobic shape, with the wavelengths of the cross-over points closely corresponding to those of the maxima in the optical absorbance spectra. These latter CD features are discussed in relation to the proposed structure of cytochrome b.
Asunto(s)
Complejo III de Transporte de Electrones/aislamiento & purificación , Mitocondrias Cardíacas/enzimología , Animales , Bovinos , Dicroismo Circular , Grupo Citocromo b/aislamiento & purificación , Citocromos c1/aislamiento & purificación , Proteínas Hierro-Azufre/aislamiento & purificación , Oxidación-ReducciónRESUMEN
N,N'-Dicyclohexylcarbodiimide (DCCD) induces a complex set of effects on the succinate-cytochrome c span of the mitochondrial respiratory chain. At concentrations below 1000 mol per mol of cytochrome c1, DCCD is able to block the proton-translocating activity associated to succinate or ubiquinol oxidation without inhibiting the steady-state redox activity of the b-c1 complex either in intact mitochondrial particles or in the isolated ubiquinol-cytochrome c reductase reconstituted in phospholipid vesicles. In parallel to this, DCCD modifies the redox responses of the endogenous cytochrome b, which becomes more rapidly reduced by succinate, and more slowly oxidized when previously reduced by substrates. At similar concentrations the inhibitor apparently stimulates the redox activity of the succinate-ubiquinone reductase. Moreover, DCCD, at concentrations about one order of magnitude higher than those blocking proton translocation, produces inactivation of the redox function of the b-c1 complex. The binding of [14C]DCCD to the isolated b-c1 complex has shown that under conditions leading to the inhibition of the proton-translocating activity of the enzyme, a subunit of about 9500 Da, namely Band VIII, is the most heavily labelled polypeptide of the complex. The possible correlations between the various effects of DCCD and its modification of the b-c1 complex are discussed.
Asunto(s)
Carbodiimidas/farmacología , Diciclohexilcarbodiimida/farmacología , Mitocondrias Cardíacas/enzimología , Mitocondrias Hepáticas/enzimología , Complejos Multienzimáticos/metabolismo , NADH NADPH Oxidorreductasas/metabolismo , Quinona Reductasas/metabolismo , Animales , Bovinos , Grupo Citocromo c/metabolismo , Complejo III de Transporte de Electrones , Cinética , Potenciales de la Membrana/efectos de los fármacos , Oxígeno/metabolismo , Ratas , Succinato Deshidrogenasa/metabolismo , Ubiquinona/metabolismoRESUMEN
We investigated the kinetics of mitochondrial ATPase in bovine heart mitochondria and submitochondrial particles upon treatment with phospholipase A2, or upon addition of n-butanol to perturb the lipid protein interactions. The changes observed are the following: (1) Lipid removal or perturbation with butanol is accompanied by loss of ATPase activity with decrease of both V and of the KM for ATP. (2) There are changes of activation energy of ATPase activity at temperatures above the discontinuity normally observed for membrane-bound enzymes in mitochondria. In particular, butanol abolishes the discontinuity, and induces a constant activation energy of about 32 kcal/mol in the range 8--37 degrees C. (3) Butanol modifies the pH dependence of ATPase shifting the pH optimum from around 10 to less alkaline values. The optimum for Mg2+ concentrations is increased by the solvent. (4) Treatment with phospholipase A2 results in a removal of oligomycin-sensitive ATPase, whereas butanol addition prevents oligomycin inhibition of ATPase. (5) In beef heart mitochondria, a spin-labelled analog of the inhibitor, dicyclohexyl carbodiimide, did not show any change in environment upon butanol addition, unlike that found in mitochondria from Saccharomyces cerevisiae.
Asunto(s)
Adenosina Trifosfatasas/metabolismo , Mitocondrias Cardíacas/enzimología , Mitocondrias/enzimología , Fosfolípidos/fisiología , Partículas Submitocóndricas/enzimología , Animales , Butanoles/farmacología , Bovinos , Cinética , Magnesio/farmacología , Mitocondrias Cardíacas/efectos de los fármacos , Oligomicinas/farmacología , Fosfolipasas , Partículas Submitocóndricas/efectos de los fármacos , TermodinámicaRESUMEN
The lipid composition and fluidity of plasma membranes have been studied at different stages of liver regeneration (4, 15 and 24 h after surgery). The phospholipid and fatty acid composition is not modified, whereas the cholesterol/phospholipid ratio is lower with respect to control membranes. The modification of the physical properties of the membranes has been studied directly by EPR analysis and indirectly by temperature dependence and cooperativity of some membrane-bound enzymes (Mg2+-ATPase, (Na+ + K+)-ATPase and 5'nucleotidase). Surgical operation or anaesthesia alone causes an early increase in fluidity; such an effect appears to be markedly reduced at a later stage. There seems to be a marked effect of regeneration on plasma membrane fluidity 15 h after partial hepatectomy when several parameters--surface fluidity, cholesterol/phospholipid ratio, and 5'-nucleotidase activity in the presence of concanavalin A -- are modified and indicate an increase in membrane fluidity. It is suggested that this modification of membrane properties could be related to the proliferative process.
Asunto(s)
Membrana Celular/metabolismo , Regeneración Hepática , Hígado/metabolismo , Fluidez de la Membrana , Lípidos de la Membrana/metabolismo , Adenosina Trifosfatasas/análisis , Animales , Membrana Celular/ultraestructura , Espectroscopía de Resonancia por Spin del Electrón , Cinética , Hígado/ultraestructura , Masculino , Nucleotidasas/análisis , Fosfolípidos/metabolismo , Ratas , TermodinámicaRESUMEN
Isolated, nucleotide-depleted bovine-heart F1-ATPase exhibits a break in Arrhenius plot with a 2.7-fold increase in activation energy of ATP hydrolysis below 18-19 degrees C. Analysis of intrinsic tyrosine fluorescence and of the circular dichroism of F1-ATPase showed an abrupt and reversible conformational change occurring at the break temperature, characteristic of a structural tightening at low temperature. Analysis of catalytic nucleotide binding sites using fluorescent ADP analog, 3'-O-(1-naphthoyl)adenosine diphosphate did not show any significant change in affinity of nucleotide binding around the transition temperature but the bound fluorophore exerted a more restricted motion and slower rotation at temperature below the break, indicating a change in the mobility of groups in the close neighbourhood. It is concluded that, as a result of temperature, two kinetically distinct states of F1-ATPase are induced, due to a change in enzyme conformation, which influences directly the properties of catalytic nucleotide binding sites.
Asunto(s)
Mitocondrias Cardíacas/enzimología , Nucleótidos/metabolismo , ATPasas de Translocación de Protón/metabolismo , Adenosina Difosfato/análogos & derivados , Adenosina Difosfato/metabolismo , Adenosina Trifosfato/metabolismo , Animales , Sitios de Unión , Catálisis , Bovinos , Dicroismo Circular , Fluorescencia , Conformación Proteica , Espectrometría de Fluorescencia , Temperatura , TermodinámicaRESUMEN
Mitochondria are strongly involved in the production of reactive oxygen species, considered as the pathogenic agent of many diseases and of aging. The mitochondrial theory of aging considers somatic mutations of mitochondrial DNA induced by oxygen radicals as the primary cause of energy decline; experimentally, complex I appears to be mostly affected and to become strongly rate limiting for electron transfer. Mitochondrial bioenergetics is also deranged in human platelets upon aging, as shown by the decreased Pasteur effect (enhancement of lactate production by respiratory chain inhibition). Cells counteract oxidative stress by antioxidants; among lipophilic antioxidants, coenzyme Q is the only one of endogenous biosynthesis. Exogenous coenzyme Q, however, protects cells from oxidative stress by conversion into its reduced antioxidant form by cellular reductases.
Asunto(s)
Envejecimiento/fisiología , Metabolismo Energético , Mitocondrias/fisiología , Animales , Antioxidantes/análisis , Antioxidantes/metabolismo , Antioxidantes/farmacología , Plaquetas/fisiología , Coenzimas , Complejo I de Transporte de Electrón , Humanos , Degeneración Macular/metabolismo , Mitocondrias/genética , Mitocondrias/metabolismo , Mitocondrias Cardíacas/metabolismo , Mitocondrias Musculares/metabolismo , NADH NADPH Oxidorreductasas/metabolismo , Estrés Oxidativo/efectos de los fármacos , Ratas , Especies Reactivas de Oxígeno/metabolismo , Ubiquinona/análogos & derivados , Ubiquinona/análisis , Ubiquinona/metabolismo , Ubiquinona/farmacologíaRESUMEN
The onset of resistance to drug-induced apoptosis of tumour cells is a major problem in cancer therapy. We studied a drug-selected clone of promyelocytic HL-60 cells, called HCW-2, which display a complex resistance to a wide variety of apoptosis-inducing agents and we found that these cells show a dramatic increase in the expression of heat shock proteins (Hsps) 70 and 27, while the parental cell line does not. It is known that stress proteins such as Hsps can confer resistance to a variety of damaging agents other than heat shock, such as TNF-alpha, monocyte-induced cytotoxicity, and also play a role in resistance to chemotherapy. This elevated expression of Hsps is paralleled by an increased activity of mitochondrial metabolism and pentose phosphate pathway, this latter leading to high levels of glucose-6-phosphate dehydrogenase and, consequently, of glutathione. Thus, the apoptotic-deficient phenotype is likely because of the presence of high levels of stress response proteins and GSH, which may confer resistance to apoptotic agents, including chemotherapy drugs. Moreover, the fact that in HCW-2 cells Hsp70 are mainly localised in mitochondria may account for the increased performances of mitochondrial metabolism. These observations could have some implications for the therapy of cancer, and for the design of combined strategies that act on antioxidant defences of the neoplastic cell.
Asunto(s)
Apoptosis , Mitocondrias/metabolismo , Oxidación-Reducción , Células Clonales , ADN Mitocondrial/análisis , Resistencia a Múltiples Medicamentos/genética , Glucosafosfato Deshidrogenasa/metabolismo , Glutatión/biosíntesis , Células HL-60 , Proteínas HSP70 de Choque Térmico/metabolismo , Humanos , Mitocondrias/ultraestructura , Vía de Pentosa Fosfato , Fenotipo , Proteínas Proto-Oncogénicas c-bcl-2/análisisRESUMEN
The complex I function in sub-mitochondrial particles was studied in platelets from patients and healthy carriers with 11778/ND4 or 3460/ND1 mtDNA point mutations associated with LHON. Both 11778/ND4 and 3460/ND1 mutations induced rotenone resistance and 11778/ND4 showed an increased K(m) for ubiquinol-2 with respect to the control group. It was concluded that even with different pathogenic mechanisms both mutations affect the quinone binding site of complex I.
Asunto(s)
Plaquetas/metabolismo , ADN Mitocondrial/genética , Mitocondrias/metabolismo , NAD(P)H Deshidrogenasa (Quinona)/metabolismo , Atrofias Ópticas Hereditarias/metabolismo , Ubiquinona/sangre , Sitios de Unión , Humanos , Cinética , Mutación Puntual , Rotenona/farmacología , Desacopladores/farmacologíaRESUMEN
In beef heart mitochondria it has been found that the Km for coenzyme Q10 of the NADH oxidation system is in the range of the membrane concentration of the quinone; this is contrary to succinate oxidation which is in Vmax with respect to quinone content. The same proportional difference between the two systems is maintained in their affinities for the exogenous acceptor CoQ1 in non-extracted mitochondria. The Km of succinate- coenzyme Q reductase for CoQ1 is reversibly lowered in CoQ-depleted mitochondria; while in contrast the Km for NADH-coenzyme Q reductase is reversibly increased by CoQ extraction. Incorporation of exogenous quinones by co-sonication with submitochondrial particles, as evidenced by fluorescence quenching of pyrene, enhances NADH-cytochrome c reductase activity in accordance with the lack of saturation of the former system.
Asunto(s)
Benzoquinonas/metabolismo , Mitocondrias Cardíacas/metabolismo , NAD(P)H Deshidrogenasa (Quinona)/metabolismo , NAD/metabolismo , Ubiquinona/metabolismo , Animales , Antioxidantes/metabolismo , Bovinos , Complejo II de Transporte de Electrones , Cinética , Complejos Multienzimáticos/metabolismo , NADH Deshidrogenasa/metabolismo , Oxidación-Reducción , Oxidorreductasas/metabolismo , Sonicación , Succinato Deshidrogenasa/metabolismoRESUMEN
The levels of coenzyme Q were determined in blood plasma and regenerating liver mitochondria of hepatectomized rats, using as controls either sham-operated or non-operated animals. Mitochondrial CoQ9 content increased in sham-operated rats, whereas it was significantly lower in hepatectomized with respect to non-operated animals. Plasma CoQ9 levels decreased dramatically in hepatectomized animals, but increased strongly in sham-operated in comparison with non-operated rats. The data suggest the possibility of a rate-limiting step in CoQ biosynthesis in hepatectomized animals.
Asunto(s)
Hepatectomía , Regeneración Hepática , Mitocondrias Hepáticas/metabolismo , Ubiquinona/metabolismo , Animales , Lipoproteínas/metabolismo , Masculino , Periodo Posoperatorio , Ratas , Ratas Wistar , Ubiquinona/sangreRESUMEN
The apparent Km for coenzyme Q10 in NADH oxidation by coenzyme Q (CoQ)-extracted beef heart mitochondria is close to their CoQ content, whereas both succinate and glycerol-3-phosphate oxidation (the latter measured in hamster brown adipose tissue mitochondria) are almost saturated at physiological CoQ concentration. Attempts to enhance NADH oxidation rate by excess CoQ incorporation in vitro were only partially successful: the reason is in the limited amount of CoQ10 that can be incorporated in monomeric form, as shown by lack of fluorescence quenching of membrane fluorescent probes; at difference with CoQ10, CoQ5 quenches probe fluorescence and likewise enhances NADH oxidation rate above normal. Attempts to enhance the CoQ content in perfused rat liver and in isolated hepatocytes failed to show uptake in the purified mitochondrial fraction. Nevertheless CoQ cellular uptake is able to protect mitochondrial activities. Incubation of hepatocytes with adriamycin induces loss of respiration and mitochondrial potential measured in whole cells by flow cytometry using rhodamine 123 as a probe: concomitant incubation with CoQ10 completely protects both respiration and potential. An experimental study of aging in the rat has shown some decrease of mitochondrial CoQ content in heart, and less in liver and skeletal muscle. In spite of the little change observed, it is reasoned that CoQ administration may be beneficial in the elderly, owing to the increased demand for antioxidants.