Acute inhibition of respiratory capacity of muscle reduces peak oxygen consumption

1990 ◽  
Vol 259 (6) ◽  
pp. C889-C896 ◽  
Author(s):  
R. M. McAllister ◽  
R. L. Terjung
Keyword(s):  
Oxygen Consumption ◽  
Electron Transport ◽  
Cytochrome C ◽  
Peak Oxygen Consumption ◽  
Dependent Manner ◽  
Cytochrome C Reductase ◽  
Energy Demands ◽  
Hindlimb Muscle ◽  
Nadh Cytochrome

Electron transport capacity of skeletal muscle was inhibited in situ in an acute dose-dependent manner with myxothiazol, a tight-binding inhibitor of ubiquinone-cytochrome c reductase, complex III of the respiratory chain. Peak oxygen consumption of rat hindlimb muscle was determined via consecutive 10-min isometric contraction (100 ms at 100 Hz) periods of increasing energy demands (4, 8, 15, 30, 45, and 60 tetani/min), using an isolated hindlimb preparation perfused with a high oxygen delivery (approximately 6-8 mumol.min-1.g-1). Peak oxygen consumption decreased from 4.61 +/- 0.19 mumol.min-1.g-1 (control) in a dose-dependent manner to 0.73 +/- 0.07 mumol.min-1.g-1 at 0.50 microM myxothiazol in blood. Oxygen extraction decreased from 65 to 12% of delivered oxygen. Furthermore, the reduction in peak respiratory rate became evident at lower energy demands of the contraction sequence. Myxothiazol inhibition of respiration was not dependent on the presence of muscle contractions but was evident when mitochondria were uncoupled with carbonyl cyanide m-chlorophenylhydrazone. A 50% effective dosage (ED50) of 0.21 microM myxothiazol for inhibition of peak oxygen consumption closely resembled the inhibition of NADH-cytochrome c reductase activity (ED50 of 0.27 microM) determined from homogenates of the same muscles. This suggests that the peak oxygen consumption of skeletal muscle is tightly coupled to the capacity for electron transport evaluated by flux through NADH-cytochrome c reductase. If the enzyme activity measured in vitro correctly represents available enzymatic capacity within contracting muscle, approximately 75% of electron transport capacity for handling reducing equivalents generated from NADH is utilized during peak oxygen consumption of rat hindlimb muscle contracting in situ.

Increased peak oxygen consumption of trained muscle requires increased electron flux capacity

1994 ◽  
Vol 77 (4) ◽  
pp. 1941-1952 ◽  
Author(s):  
D. M. Robinson ◽  
R. W. Ogilvie ◽  
P. C. Tullson ◽  
R. L. Terjung
Keyword(s):  
Electron Transport ◽  
Cytochrome C ◽  
Reductase Activity ◽  
Tight Binding ◽  
Treadmill Running ◽  
Complex Iii ◽  
Peak Oxygen Consumption ◽  
Transport Capacity ◽  
Cytochrome C Reductase ◽  
Nadh Cytochrome

The importance of the training-induced increase in mitochondrial capacity in realizing the increase in maximal O2 consumption (VO2max) of trained muscle was evaluated using an isolated perfused rat hindlimb preparation at a high blood flow (approximately 80 ml.min-1.100 g-1) during tetanic contractions. Rats trained for 8-–12 wk by treadmill running exhibited an approximately 25% increase in muscle VO2max (5.62 +/- 0.31 to 7.06 +/- 0.64 mumol.min-1.g-1), an increase in mitochondrial enzyme activity (approximately 70% for cytochrome oxidase and approximately 55% for NADH cytochrome-c reductase), and an increase in tissue capillarity (14%) that is expected to increase the O2 exchange capacity of the tissue. Muscle VO2max of sedentary (n = 34) and trained (n = 30) animals was determined, and electron transport capacity was acutely managed with myxothiazol, a tight-binding inhibitor of complex III. Inhibition of complex III was similar among 1) the low- and high-oxidative fibers and 2) the superficial and deep mitochondrial populations within muscle. Inhibition of NADH cytochrome-c reductase activity resulted in reductions in muscle VO2max with similar dose responses (mean effective dose of approximately 0.2 microM) of myxothiazol added to the perfusion medium. The extraction of O2 by the contracting muscle decreased as VO2max declined. The increase in muscle VO2max observed in the muscle of trained animals was eliminated when its electron transport capacity was reduced to that observed in normal sedentary rat muscle. Thus, the exercise-induced adaptation of an increased muscle mitochondrial content appears to be essential for trained muscle to exhibit its increased O2 flux capacity. The results of the present experiment illustrate the importance of mitochondrial adaptations in muscle remodeled by exercise training.

Endoplasmic reticulum membrane isolated from small-intestinal epithelial cells: enzyme and protein components

1981 ◽  
Vol 52 (1) ◽  
pp. 215-222
Author(s):  
M. Fujita ◽  
H. Ohta ◽  
T. Uezato
Keyword(s):  
Endoplasmic Reticulum ◽  
Epithelial Cells ◽  
Cytochrome C ◽  
Endoplasmic Reticulum Membrane ◽  
Dependent Manner ◽  
Cytochrome C Reductase ◽  
Basolateral Membranes ◽  
Nadh Cytochrome ◽  
Protein Components ◽  
A Minor

Endoplasmic reticulum membrane-rich fraction was obtained by subfractionation of the light microsomes from mouse jejunal mucosal epithelial cells. It was marked by high glucose-6-phosphatase, NADPH-cytochrome c reductase, and NADH-cytochrome c reductase activities and low Na+,K+-ATPase activity. The enrichment of Na+,K+-ATPase was 180-fold higher in the basolateral membranes than in the endoplasmic reticulum membrane-rich fraction relative to glucose-6-phosphatase. The protein peak that was phosphorylated in a Na-dependent manner was prominent in the basolateral membranes while it was a minor peak in the endoplasmic reticulum membrane-rich fraction. Under the electron microscope the fraction was seen to be composed of homogeneous small vesicles with thin smooth membranes.

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.

scholarly journals AN ELECTRON-TRANSPORT SYSTEM ASSOCIATED WITH THE OUTER MEMBRANE OF LIVER MITOCHONDRIA

1967 ◽  
Vol 32 (2) ◽  
pp. 415-438 ◽  
Author(s):  
Gian Luigi Sottocasa ◽  
Bo Kuylenstierna ◽  
Lars Ernster ◽  
Anders Bergstrand
Keyword(s):  
Electron Transport ◽  
Cytochrome C ◽  
Outer Membrane ◽  
Respiratory Chain ◽  
Transport System ◽  
Cytochrome B5 ◽  
Liver Mitochondria ◽  
Electron Transport System ◽  
Cytochrome C Reductase ◽  
Nadh Cytochrome

Preparations of rat-liver mitochondria catalyze the oxidation of exogenous NADH by added cytochrome c or ferricyanide by a reaction that is insensitive to the respiratory chain inhibitors, antimycin A, amytal, and rotenone, and is not coupled to phosphorylation. Experiments with tritiated NADH are described which demonstrate that this "external" pathway of NADH oxidation resembles stereochemically the NADH-cytochrome c reductase system of liver microsomes, and differs from the respiratory chain-linked NADH dehydrogenase. Enzyme distributation data are presented which substantiate the conclusion that microsomal contamination cannot account for the rotenone-insensitive NADH-cytochrome c reductase activity observed with the mitochondria. A procedure is developed, based on swelling and shrinking of the mitochondria followed by sonication and density gradient centrifugation, which permits the separation of two particulate subfractions, one containing the bulk of the respiratory chain components, and the other the bulk of the rotenone-insensitive NADH-cytochrome c reductase system. Morphological evidence supports the conclusion that the former subfraction consists of mitochondria devoid of outer membrane, and that the latter represents derivatives of the outer membrane. The data indicate that the electron-transport system associated with the mitochondrial outer membrane involves catalytic components similar to, or identical with, the microsomal NADH-cytochrome b5 reductase and cytochrome b5.

scholarly journals Changes in Microsomal Electron Transport of Plant Storage Tissues Induced by Slicing and Aging

1972 ◽  
Vol 25 (1) ◽  
pp. 103 ◽  
Author(s):  
JM Rungie ◽  
JT Wiskich
Keyword(s):  
Endoplasmic Reticulum ◽  
Electron Transport ◽  
Cytochrome C ◽  
Nadh Dehydrogenase ◽  
Microsomal Protein ◽  
Partial Recovery ◽  
Cytochrome C Reductase ◽  
Nadh Cytochrome ◽  
Induced Changes ◽  
Plant Storage

Slicing turnip, swede, and beet storage tissues induced 20-100% loss of micro-somal NADH dehydrogenase activities within 10 min. Subsequent washing of the slices resulted in partial recovery of some activities particularly NADH-cytochrome c reductase which reached a maximum after 24 hr aging then again declined. Slicing also induced a 20% decrease in microsomal protein but this loss was recovered after 5-10 hr aging. These induced changes correlated with reported changes in the ultra-structure of the endoplasmic reticulum.

scholarly journals Properties of a cytochrome c-enriched light particulate fraction isolated from the photosynthetic bacterium Rhodopseudomonas spheroides

1978 ◽  
Vol 174 (1) ◽  
pp. 267-275 ◽  
Author(s):  
J Barrett ◽  
C N Hunter ◽  
O T G Jones
Keyword(s):  
Cytochrome C ◽  
Sodium Dodecyl ◽  
Photosynthetic Bacterium ◽  
Particulate Fraction ◽  
Cytochrome C Reductase ◽  
Cytochrome C2 ◽  
Succinate Cytochrome C Reductase ◽  
Bulk Membrane ◽  
Nadh Cytochrome ◽  
Rhodopseudomonas Spheroides

Differential centrifugation of suspensions of French-press-disrupted Rhodopseudomonas spheroides yielded a light particulate fraction that was different in many properties from the bulk membrane fraction. It was enriched in cytochrome c and had a low cytochrome b content. When prepared from photosynthetically grown cells this fraction had a very low specific bacteriochlorophyll content. The cytochrome c of the light particles differed in absorption maxima at 77K from cytochrome c2 attached to membranes; there was pronounced splitting of the alpha-band, as is found in cytochrome c2 free in solution. Potentiometric titration at A552–A540 showed the presence of two components that fitted an n = 1 titration; one component had a midpoint redox potential of +345mV, like cytochrome c2 in solution, and the second had E0′ at pH 7.0 of +110 mV, and they were present in a ratio of approx. 2:3. Difference spectroscopy at 77K showed that the spectra of the two components were very similar. More of a CO-binding component was present in particles from photosynthetically grown cells. Light membranes purified by centrifugation on gradients of 5–60% (w/w) sucrose retained the two c cytochromes; they contained no detectable succinate-cytochrome c reductase or bacteriochlorophyll and very little ubiquinone, but they contained NADH-cytochrome c reductase and some phosphate. Electrophoresis on sodium dodecyl sulphate/polyacrylamide gels showed that the light membranes of aerobically and photosynthetically grown cells were very similar and differed greatly from other membrane fractions of R. spheroides.

Ferrocyanide-activated NADH cytochrome c reductase from barley

1980 ◽  
Vol 18 (4) ◽  
pp. 389-393 ◽  
Author(s):  
Ian S. Small ◽  
John L. Wray
Keyword(s):  
Cytochrome C ◽  
Cytochrome C Reductase ◽  
Nadh Cytochrome

Electron Transport and Coupled Energy Generation in Pseudomonas saccharophila

1971 ◽  
Vol 49 (11) ◽  
pp. 1175-1182 ◽  
Author(s):  
M. Ishaque ◽  
A. Donawa ◽  
M. I. H. Aleem
Keyword(s):  
Oxygen Consumption ◽  
Electron Transport ◽  
Cytochrome C ◽  
Atp Synthesis ◽  
Succinate Oxidation ◽  
Atp Formation ◽  
Low Concentrations ◽  
Oxidase Enzyme ◽  
Succinate Oxidase ◽  
Pseudomonas Saccharophila

The respiratory chain system of heterotrophically grown Pseudomonas saccharophila contained cytochromes of the b, c, a, and o types and also the NADH and succinate oxidase enzyme systems. Cell-free extracts catalyzed phosphorylation coupled to the oxidation of NADH, succinate, and ascorbate (plus cytochrome c). The P/O ratios were in the range of 1.00 for generated NADH, 0.29 for added NADH, 0.50 for succinate, and 0.25 for ascorbate (plus cytochrome c).The oxidative phosphorylation was uncoupled by 2,4-dinitrophenol, 2,6-dibromophenol, pentachlorophenol, m-chlorocarbonyl cyanide phenylhydrazone, and dicumarol without any inhibition of oxygen consumption. Phosphorylation coupled to NADH oxidation was completely inhibited by the flavoprotein inhibitors such as rotenone, amytal, and atabrine; these inhibitors had no effect, however, on the ATP synthesis associated with succinate oxidation. Antimycin A or 2-n-nonyl-4-hydroxyquinoline-N-oxide as well as cyanide or azide at low concentrations completely inhibited the phosphate esterification coupled to the oxidation of NADH or succinate, but had little or no effect on the oxygen consumption. Relatively higher concentrations of oligomycin were required for a complete inhibition of the electron-transport-linked ATP formation.

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