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1.
Physical exercise and fatty acids have been studied in relation to mitochondrial composition and function in rat liver, heart, and skeletal muscle. Male rats were divided into two groups according to dietary fat type (virgin olive and sunflower oils). One-half of the animals from each group were subjected to a submaximal exercise for 8 weeks; the other half acted as sedentary controls. Coenzyme Q, cytochromes b, c + c1, a + a3 concentrations, and the activity of cytochrome c oxidase were determined. Regular exercise increased (P < 0.05) the concentration of the above-mentioned elements and the activity of the cytochrome c oxidase by roughly 50% in liver and skeletal muscle. In contrast, physical exercise decreased (P < 0.05) cytochrome c oxidase activity in the heart (in micromol/min/g, from 8.4+/-0.1 to 4.9+/-0.1 in virgin olive oil group and from 9.7+/-0.1 to 6.7+/-0.2 in sunflower oil animals). Dietary fat type raised the levels of coenzyme Q, cytochromes, and cytochrome c oxidase activity in skeletal muscle (P < 0.05) among the rats fed sunflower oil. In conclusion, dietary fat type, regular exercise, and the specific tissue modulate composition and function of rat mitochondria.  相似文献   

2.
We investigate mitochondrial-lipid peroxidation of mitotic (liver) and postmitotic (heart and skeletal muscle) tissues of rats fed lifelong on two different lipid sources: virgin olive oil (monounsaturated fatty acids) and sunflower oil (n–6 polyunsaturated fatty acids). Two groups of 80 rats each were fed over 24 months on a diet differing in the lipid source (virgin olive oil or sunflower oil). Twenty rats per group were killed at 6, 12, 18, and 24 months; liver, heart, and skeletal muscle mitochondria were isolated and the lipid profile, hydroperoxides, vitamin E, and ubiquinone as well as catalase activity measured. Lipid peroxidation was higher in postmitotic tissues, and sunflower oil led to a higher degree of polyunsaturation and peroxidation. The levels of -tocopherol adapted to oxidative stress and preferentially accumulated during aging in heart and skeletal muscle. In conclusion, the type of dietary fat should be considered in studies on aging, since oxidative stress is directly modulated by this factor. This study confirms that postmitotic tissues are more prone to oxidative stress during aging and proposes a hypothesis to explain this phenomenon.  相似文献   

3.
Fat frying is a popular food preparation method but several components like antioxidant vitamins could be lost due to oxidation and some others with toxic effects could appear. Because of such large consumption of frying oils, the effect of high temperatures on the oils is of major concern both for product quality and nutrition, taking into account that dietary fat source deeply influences several biochemical parameters, especially of mitochondrial membranes. Virgin olive oil possesses specific features for modulating the damages occurred by endogenous and exogenous oxidative stress being particularly rich in antioxidant molecules. We evaluated the extent of modifications suffered by virgin olive oil following a short-time deep fat frying procedure: vitamin E and phenolic compound as well as total antioxidant capacity (measured by ESR) decreased, while polar compounds increased. The intake of such an altered oil mainly affected the hydroperoxide and TBARS contents of mitochondrial membranes which were enhanced after the dietary treatments. Also, several mitochondrial respiratory chain components (Coenzyme Q, cytochrome b, c + c 1, and a + a 3) were affected.  相似文献   

4.
Summary Cold acclimation lowers the selected body temperature (T b) in many ectothermic vertebrates. This change in behavioural thermoregulation is accompanied by an increase in the proportion of polyunsaturated fatty acids in tissues and cellular membranes. We investigated how diets containing different fatty acids, known to significantly alter the fatty acid composition of animal tissues and membranes, affect the selected T b of the lizard Tiliqua rugosa. Lizards on a diet containing many polyunsaturated fatty acids (10% sunflower oil) showed a 3–5°C decrease in T b, whereas T b in animals on a diet containing mainly saturated fatty acids (10% sheep fat) did not change. Our study suggests that the composition of dietary lipids influences thermoregulation in ectothermic vertebrates and may thus play a role in the seasonal adjustment of their physiology.Abbreviations CST central standard time - T a air temperature - T b Body temperature  相似文献   

5.
In this study we have evaluated the supplementation of olive oil with vitamin E on coenzyme Q concentration and lipid peroxidation in rat liver mitochondrial membranes. Four groups of rats were fed on virgin olive, olive plus 200 mg/kg of vitamin E or sunflower oils as lipid dietary source. To provoke an oxidative stress rats were administered intraperitoneally 10 mg/kg/day of adriamycin the last two days of the experiment. Animals fed on olive oil plus vitamin E had significantly higher coenzyme Q and vitamin E levels but a lower mitochondrial hydroperoxide concentration than rats fed on olive oil. Retinol levels were not affected, by either different diets or adriamycin treatment. In conclusion, an increase in coenzyme Q and alpha-tocopherol in these membranes can be a basis for protection against oxidation and improvement in antioxidant capacity.  相似文献   

6.
Mitochondria-related oxidative damage is a primary event in aging and age-related neurodegenerative disorders. Some dietary treatments, such as antioxidant supplementation or the enrichment of mitochondrial membranes with less oxidizable fatty acids, reduce lipid peroxidation and lengthen life span in rodents. This study compares life-long feeding on monounsaturated fatty acids (MUFAs), such as virgin olive oil, and n-6 polyunsaturated fatty acids, such as sunflower oil, with or without coenzyme Q10 supplementation, with respect to age-related molecular changes in rat brain mitochondria. The MUFA diet led to diminished age-related phenotypic changes, with lipoxidation-derived protein markers being higher among the older animals, whereas protein carbonyl compounds were lower. It is noteworthy that the MUFA diet prevented the age-related increase in levels of mitochondrial DNA deletions in the brain mitochondria from aged animals. The findings of this study suggest that age-related oxidative stress is related, at the mitochondrial level, to other age-related features such as mitochondrial electron transport and mtDNA alterations, and it can be modulated by selecting an appropriate dietary fat type and/or by suitable supplementation with low levels of the antioxidant/electron carrier molecule coenzyme Q.  相似文献   

7.
We investigated how dietary fats and oils of different fatty acid composition influence the seasonal change of body mass, fur colour, testes size and torpor in Djungarian hamsters, Phodopus sungorus, maintained from autumn to winter under different photoperiods and temperature regimes. Dietary fatty acids influenced the occurrence of spontaneous torpor (food and water ad libitum) in P. sungorus maintained at 18°C under natural and artificial short photoperiods. Torpor was most pronounced in individuals on a diet containing 10% safflower oil (rich in polyunsaturated fatty acids), intermediate in individuals on a diet containing 10% olive oil (rich in monounsaturated fatty acids) and least pronounced in individuals on a diet containing 10% coconut fat (rich in saturated fatty acids). Torpor in P. sungorus on chow containing no added fat or oil was intermediate between those on coconut fat and olive oil. Dietary fatty acids had little effect on torpor in animals maintained at 23°C. Body mass, fur colour and testes size were also little affected by dietary fatty acids. The fatty acid composition of brown fat from hamsters maintained at 18°C and under natural photoperiod strongly reflected that of the dietary fatty acids. Our study suggests that the seasonal change of body mass, fur colour and testes size are not significantly affected by dietary fatty acids. However, dietary fats influence the occurrence of torpor in individuals maintained at low temperatures and that have been photoperiodically primed for the display of torpor.Abbreviations BAT brown adipose tissue - bm body mass - FA fatty acid(s) - MR metabolic rate - MUFA monounsaturated fatty acid(s) - PUFA polyunsaturated fatty acid(s) - SFA saturated fatty acid(s) - T a air temperature - T b body temperature - Ts body surface temperature(s) - TNZ thermoneutral zone - UFA unsaturated fatty acid(s)  相似文献   

8.
The mitochondrial theory of aging suggests that this phenomenon is the consequence of random somatic mutations in mitochondrial DNA, induced by long-term exposure to free radical attack. There are two potential dietary means of delaying the effects of free radicals on cellular aging, i.e., enrichment of mitochondrial membranes with monounsaturated fatty acids and supplementation with antioxidants. We have performed a preliminary study on male rats, 6 or 12 month old, fed with diets differing in the nature of the fat (virgin olive oil or sunflower oil) and/or with antioxidant supplementation (coenzyme Q10), analysing hydroperoxide and coenzyme Q9 and Q10 in heart mitochondria. Preliminary results allow us to conclude that the CoQ10 dietetic supplementation as well as the enrichment of the cellular membranes with monounsaturated fatty acids, successfully protect mitochondrial membranes from aged rats against the free radical insult.  相似文献   

9.
The effects of five different dietary fat supplements on fatty acid composition and oxidative stability of subcutaneous and kidney fat were evaluated in 36 Brown Swiss bulls and compared to a low fat diet in a monofactorial design. The following fat supplements were provided as additional fat at 30 g per kg feed dry matter: crystalline rumen‐protected fat, coconut oil, and three types of crushed whole oilseeds (rapeseed, sunflower seed and linseed). Adipose tissues reflected differences (P < 0.05) in dietary fatty acid composition although to a lower extent. Using protected fat, which contained elevated levels of trans fatty acids, and sunflower seed, containing a high proportion of linoleic acid, significantly increased C18:1 trans fatty acid proportion in the adipose tissues. The use of sunflower seed increased conjugated linoleic acid. The oilseeds resulted in lower amounts of C16:0 in favour of C18:0. Except for linseed, all fat supplemented groups improved oxidative stability of adipose tissues as compared with control. This was explained by lower proportions of unsaturated fatty acids in adipose tissue (protected fat), by elevated α‐tocopherol contents (rapeseed, sunflower seed) or by a combination of both (coconut oil). Fat colour remained unaffected by treatments. Compared to other fat supplements oilseeds, especially sunflower seed and rapeseed, can therefore be recommended to be fed to bulls in order to increase the proportions of C18 unsaturated fatty acids in adipose tissues and to maintain or improve oxidative stability.  相似文献   

10.
Summary.  Wistar rats were fed with different diets with or without supplement coenzyme Q10 (CoQ10) and with oil of different sources (sunflower or virgin olive oil) for six or twelve months. Ubiquinone contents (CoQ9 and CoQ10) were quantified in homogenates of livers and brains from rats fed with the four diets. In the brain, younger rats showed a 3-fold higher amount of ubiquinone than older ones for all diets. In the liver, however, CoQ10 supplementation increased the amount of CoQ9 and CoQ10 in both total homogenates and plasma membranes. Rats fed with sunflower oil as fat source showed higher amounts of ubiquinone content than those fed with olive oil, in total liver homogenates, but the total ubiquinone content in plasma membranes was similar with both fat sources. Older rats showed a higher amount of ubiquinone after diets supplemented with CoQ10. Two ubiquinone-dependent antioxidant enzyme activities were measured. NADH-ferricyanide reductase activity in hepatocyte plasma membranes was unaltered by ubiquinone accumulation, but this activity increased slightly with age. Both cytosolic and membrane-bound dicumarol-sensitive NAD(P)H:(quinone acceptor) oxidoreductase (DT-diaphorase, EC 1.6.99.2) activities were decreased by diets supplemented with CoQ10. Animals fed with olive oil presented lower DT-diaphorase activity than those fed with sunflower oil, suggesting that the CoQ10 antioxidant protection is strengthened by olive oil as fat source. Received May 22, 2002; accepted September 20, 2002; published online May 21, 2003 RID="*" ID="*" Correspondence and reprints: Departamento de Biología Celular, Fisiología e Inmunología, Universidad de Córdoba Edificio Severo Ochoa, Campus de Rabanales, 14071 Córdoba, Spain.  相似文献   

11.
12.
The triphasic course previously reported for the reduction of cytochrome b in the succinate-cytochrome c reductase by either succinate or duroquinol has been shown to be dependent on the redox state of the enzyme preparation. Prior reduction with increasing concentrations of ascorbate leads to partial reduction of cytochrome c1, and a gradual decrease in the magnitude of the oxidation phase of cytochrome b. At an ascorbate concentration sufficient to reduce cytochrome c1 almost completely, the reduction of cytochrome b by either succinate or duroquinol becomes monophasic. Owing to the presence of a trace amount of cytochrome oxidase in the reductase preparation employed, the addition of cytochrome c makes electron flow from substrate to oxygen possible. Under such circumstances, the addition of a limited amount of either succinate or duroquinol leads to a multiphasic reduction and oxidation of cytochrome b. After the initial three phases as described previously, cytochrome b becomes oxidized before cytochrome c1 when the limited amount of added substrate is being used up. However, at the end of the reaction when cytochrome ca is being rapidly oxidized, cytochrome b becomes again reduced. The above observations support a cyclic scheme of electron flow in which the reduction of cytochrome b proceeds by two different routes and its oxidation controlled by the redox state of a component of the respiratory chain.  相似文献   

13.
In this work, we investigated whether cold exposure-induced hyperthyroidism increases oxidative damage and susceptibility to oxidants of rat liver, heart and skeletal muscle. All tissues exhibited gradual increases in hydroperoxide and protein-bound carbonyl levels. Glutathione peroxidase activity increased in all tissues after 2 days and further increased in the muscle after 10 days of cold exposure. Liver glutathione reductase activity increased after 10 days of cold exposure, while heart and muscle activities were not modified. Vitamin E levels were not affected by cold, while coenzyme Q9 and coenzyme Q10 levels decreased in heart and muscle after 2-day cold exposure and were not further modified after 10 days. Liver coenzyme Q9 levels increased after 2 days whereas coenzyme Q10 levels increased after 10 days in the cold. The whole antioxidant capacity was lowered, while parameters positively correlated with susceptibility to oxidants were increased by cold. Lipid fatty acid composition was modified in all tissues. In particular, fatty acid unsaturation degree increased in heart and muscle. Cytochrome oxidase activity increased, suggesting an increased content of hemoproteins, which are able to generate .OH radical. This view was supported by the observation that the tissue susceptibility to H(2)O(2) treatment, which is strongly correlated to iron-ligand content, increased after cold exposure. In this frame, it is apparent that the increase in oxidative capacity, necessary for homeotherm survival in low temperature environments, has potential harmful effects, because it results in increased susceptibility to oxidative challenge.  相似文献   

14.
The protective effects of dietary pyrroloquinoline quinone disodium (PQQ.Na2) supplementation against oxidized sunflower oil-induced oxidative stress and liver injury in laying hens were examined. Three hundred and sixty 53-week-old Hy-Line Gray laying hens were randomly allocated into one of the five dietary treatments. The treatments included: (1) a diet containing 2% fresh sunflower oil; (2) a diet containing 2% thermally oxidized sunflower oil; (3) an oxidized sunflower oil diet with 100 mg/kg of added vitamin E; (4) an oxidized sunflower oil diet with 0.08 mg/kg of PQQ.Na2; and (5) an oxidized sunflower oil diet with 0.12 mg/kg of PQQ.Na2. Birds fed the oxidized sunflower oil diet showed a lower feed intake compared to birds fed the fresh oil diet or oxidized oil diet supplemented with vitamin E (P=0.009). Exposure to oxidized sunflower oil increased plasma malondialdehyde (P<0.001), hepatic reactive oxygen species (P<0.05) and carbonyl group levels (P<0.001), but decreased plasma glutathione levels (P=0.006) in laying hens. These unfavorable changes induced by the oxidized sunflower oil diet were modulated by dietary vitamin E or PQQ.Na2 supplementation to levels comparable to the fresh oil group. Dietary supplementation with PQQ.Na2 or vitamin E increased the activities of total superoxide dismutase and glutathione peroxidase in plasma and the liver, when compared with the oxidized sunflower oil group (P<0.05). PQQ.Na2 or vitamin E diminished the oxidized sunflower oil diet induced elevation of liver weight (P=0.026), liver to BW ratio (P=0.001) and plasma activities of alanine aminotransferase (P=0.001) and aspartate aminotransferase (P<0.001) and maintained these indices at the similar levels to the fresh oil diet. Furthermore, oxidized sunflower oil increased hepatic DNA tail length (P<0.05) and tail moment (P<0.05) compared with the fresh oil group. Dietary supplementation of PQQ.Na2 or vitamin E decreased the oxidized oil diet induced DNA tail length and tail moment to the basal levels in fresh oil diet. These results indicate that PQQ.Na2 is a potential antioxidant and is as effective against oxidized oil-related liver injury in laying hens as vitamin E. The protective effects of PQQ.Na2 against liver damage induced by oxidized oil may be partially due to its role in the scavenging of free radicals, inhibiting of lipid peroxidation and enhancing of antioxidant defense systems.  相似文献   

15.
Thiobacillus tepidarius was shown to contain cytochrome(s) c with absorption maxima at 421, 522 and 552 nm in room temperature reduced minus oxidized difference spectra, present at 1.1–1.2 nmol per mg dry wt and present in both membrane and soluble fractions of the cell. The membrane-bound cytochrome c (1.75 nmol per mg membrane protein) had a midpoint potential (Em, pH 7.0) of 337 mV, while the soluble fractions appeared to contain cytochrome(s) c with Em (pH 7.0) values of about 270 and 360 mV. The organism also contained three distinct membrane-bound b-type cytochromes (totalling 0.33 nmol per mg membrane protein), each with absorption maxima in reduced minus oxidized difference spectra at about 428, 532 and 561 nm. The Em (pH 7.0) values for the three cytochromes b were 8 mV (47.8% of total), 182 mV (13.7%) and 322 mV (38.5%). No a- or d-type cytochromes were detectable spectrophotometrically in the intact organism or its membrane and soluble fractions. Evidence is presented for both CO-binding and CO-unreactive cytochromes b or o, and CO-binding cytochrome(s) c. From redox effects observed with CO it is proposed that a cytochrome c donates electrons to a cytochrome b, and that a high potential cytochrome b or o may be acting as the terminal oxidase in substrate oxidation. This may be the 445 nm pigment, a photodissociable CO-binding membrane haemoprotein. Substrate oxidation was relatively insensitive to CO-inhibition, but strongly inhibited by cyanide and azide. Thiosulphate oxidation couples directly to cytochrome c reduction, but tetrathionate oxidation is linked (probably via ubiquinone Q-8) to reduction of a cytochrome b of lower potential than the cytochrome c. The nature of possible electron transport pathways in Thiobacillus tepidarius is discussed. One speculative sequence is: c b8 b182 c270 c337 b322/c360 O2 Abbreviations Em midpoint electrode potential - E inf0 sup pH 7, standard electrode potential at pH 7.0 - Q-8 coenzyme Q-8 (ubiquinone-40)  相似文献   

16.
Currently, diets higher in polyunsaturated fat are believed to lower blood cholesterol concentrations, and thus reduce atherosclerosis, greater than diets containing high amounts of saturated or possibly even monounsaturated fat. The present study was designed to investigate the effect of diets containing mid- or high-linoleic oil versus the typical high-linoleic sunflower oil on LDL oxidation and the development of early atherosclerosis in a hypercholesterolemic hamster model. Animals were fed a hypercholesterolemic diet containing 10% mid-oleic sunflower oil, high-oleic olive oil, or high-linoleic sunflower oil (wt/wt) plus 0.4% cholesterol (wt/wt) for 10 weeks. After 10 weeks of dietary treatment, only the animals fed the mid-oleic sunflower oil had significant reductions in plasma LDL-C levels (-17%) compared to the high-linoleic sunflower oil group. The high-oleic olive oil-fed hamsters had significantly higher plasma triglyceride levels (+41%) compared to the high-linoleic sunflower oil-fed hamsters. The tocopherol levels in plasma LDL were significantly higher in hamsters fed the mid-oleic sunflower oil (+77%) compared to hamsters fed either the high-linoleic sunflower or high-oleic olive oil. Measurements of LDL oxidation parameters, indicated that hamsters fed the mid-oleic sunflower oil and high-oleic olive oil diets had significantly longer lag phase (+66% and +145%, respectively) and significantly lower propagation rates (-26% and -44%, respectively) and conjugated dienes formed (-17% and -25%, respectively) compared to the hamsters fed the high-linoleic sunflower oil. Relative to the high-linoleic sunflower oil, aortic cholesterol ester was reduced by -14% and -34% in the mid-oleic sunflower oil and high-oleic olive oil groups, respectively, with the latter reaching statistical significance. Although there were no significant associations between plasma lipids and lipoprotein cholesterol with aortic total cholesterol and cholesterol esters for any of the groups, the lag phase of conjugated diene formation was inversely associated with both aortic total and esterified cholesterol in the high-oleic olive oil-fed hamsters (r = -0.69, P < 0.05). The present study suggests that mid-oleic sunflower oil reduces risk factors such as lipoprotein cholesterol and oxidative stress associated with early atherosclerosis greater than the typical high-linoleic sunflower oil in hypercholesterolemic hamsters. The high-oleic olive oil not only significantly reduced oxidative stress but also reduced aortic cholesterol ester, a hallmark of early aortic atherosclerosis greater than the typical high-linoleic sunflower oil.  相似文献   

17.
This work was designed to study the effect of different lipid sources on the activities of lipoprotein lipase and lipogenic enzymes in adipose tissue from rats fedad libitum or energy-controlled diets. Male Wistar rats were fed diets containing 40% of energy as fat (olive oil, sunflower oil, palm oil or beef tallow), for 4 wk. Underad libitum feeding no differences were found among dietary fat groups in final body weight, adipose tissue weights and total body fat. Under energy-controlled feeding, despite isoenergetic intake, rats fed the beef tallow diet gained significantly less weight than rats fed the other three diets. Beef tallow fed rats showed the lowest values for adipose tissue weights and total body fat. When rats had free access to food no effect of dietary lipid source on lipogenic enzyme activities was found. In contrast, under energy-controlled feeding rats fed the beef tallow diet showed significantly higher activities of glucose-6-phosphate dehydrogenase and fatty acid synthase than rats fed the other three diets. Heparin-releasable lipoprotein lipase activity in perirenal and subcutaneous adipose tissues was not different among rats fed olive oil, safflower oil, palm oil or beef tallow. When comparing both adipose tissue anatomical locations, significantly higher activities were found in subcutaneous than in perirenal fat pad independently of dietary fat. In conclusion, under our experimental protocol, lipogenesis in rat adipose tissue does not seem to be affected by dietary fat type.  相似文献   

18.
The composition of tissue and membrane fatty acids in ectothermic vertebrates is influenced by both temperature acclimation and diets. If such change in body lipid composition and thermal physiology were linked, a diet-induced change in body lipid composition should result in a change in thermal physiology. We therefore investigated whether the selected body temperature of the agamid lizardAmphibolurus nuchalis (body mass 20 g) is influenced by the lipid composition of dietary fatty acids and whether diet-induced changes in thermal physiology are correlated with changes in body lipid composition. The selected body temperature in two groups of lizards was indistinguishable before dietary treatments. The selected body temperature in lizards after 3 weeks on a diet rich in saturated fatty acids rose by 2.1 °C (photophase) and 3.3 °C (scotophase), whereas the body temperature of lizards on a diet rich in unsaturated fatty acids fell by 1.5 °C (photophase) and 2.0 °C (scotophase). Significant diet-induced differences were observed in the fatty acid composition of depot fat, liver and muscle. These observations suggest that dietary lipids may influence selection of body temperature in ectotherms via alterations of body lipid composition.Abbreviations bm body mass - FA fatty acid(s) - MUFA monounsaturated fatty acids - PUFA polyunsaturated fatty acids - SFA saturated fatty acids - T a air temperature - T b body temperature - UFA unsaturated fatty acids  相似文献   

19.
Lactobacilli isolated from the rumen of cattle were subjected to morphological and biochemical characterizations followed by PCR-based identification. Among isolates, Lactobacillus brevis was found to be the most prevalent species in the rumen. For in vitro conjugated linoleic acid (CLA) production, the two isolates of L. brevis and one each of Lactobacillus viridescens and Lactobacillus lactis were selected. The sunflower oil (i.e., 0.25, 0.5 and 1.0%; a rich source of linoleic acid) was added to skim milk as a substrate for CLA production by isolates at 37 °C/12 h. L. brevis 02 was found to be the most potential CLA producer (10.53 mg CLA/g fat) at 0.25% concentration of sunflower oil followed by L. brevis 01 (8.27 mg CLA/g fat). However, at higher level of sunflower oil (i.e., 1.0%), L. lactis was the highest CLA producer (9.22 mg/g fat) when compared to L. brevis and L. viridescens. The results indicated that L. brevis and/or CLA production was inhibited with increasing concentration of sunflower oil in skim milk. In contrast, L. lactis and L. viridescens could tolerate the increasing concentrations of sunflower oil and produced higher CLA. Overall, L. brevis extends a possibility to be used as a direct-fed microbial for ruminants to increase the CLA content in milk, however, in vivo trials are needed for validation of results obtained.  相似文献   

20.
Mitochondria (or mitoplasts) and submitochondrial particles from yeast were treated with [125I] diazobenzenesulfonate to label selectively proteins exposed on the outer or inner surface of the inner mitochondrial membrane. Polyacrylamide gel analysis of the immunoprecipitates formed with antibodies against Complex III or cytochromeb revealed that the two core proteins and cytochromeb were labeled in both mitochondria and submitochondrial particles, suggesting that these proteins span the membrane. Cytochromec 1 and the iron sulfur protein were labeled in mitochondria but not in submitochondrial particles, suggesting that these proteins are exposed on the cytosolic side of the inner membrane. The steady-state reduction of cytochromesb andc 1 was determined with succinate and the decyl analogue of coenzyme Q as substrates. Addition of the coenzyme Q analogue to mitochondria caused reduction of 15–30% of the total dithionite-reducibleb and 100% of the cytochromec 1: Addition of the coenzyme Q analogue to submitochondrial particles led to the reduction of 70% of the total dithionite-reducible cytochromeb but insignificant amounts of cytochromec 1. A model to explain the topography of Complex III in the inner membrane is proposed based on these results.Abbreviations used: DABS, diazobenzene sulfonate; DBH2, reduced form of decyl analogue of coenzyme Q (2,3-dimethoxy-5-methyl-6-n-decyl-1,4-benzoquinone); PMSF, phenylmethylsulfonyl fluoride; SDS, sodium dodecyl sulfate.  相似文献   

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