PIGEON MUSCLE MITOCHONDRIA: ION CONCENTRATIONS, OXIDATIVE PHOSPHORYLATION, SWELLING, AND EXTRAPARTICULATE SPACE

1963 ◽  
Vol 41 (1) ◽  
pp. 2529-2542 ◽  
Author(s):  
G. J. Marcus ◽  
J. F. Manery
Keyword(s):  
Oxygen Consumption ◽  
Oxidative Phosphorylation ◽  
Respiratory Control ◽  
Sodium Potassium ◽  
Density Measurements ◽  
Muscle Mitochondria ◽  
Ion Concentrations ◽  
Heat Stable ◽  
Control Respiration ◽  
Mitochondrial Volume

Metabolically active mitochondria were isolated from pigeon breast muscle in cold 0.45 M sucrose with ATP, EDTA, and tris buffer at pH 7.4. Fresh mitochondria were analyzed for sodium, potassium, nitrogen, iron, and water. They were morphologically intact and showed respiratory control, respiration being limited by the phosphate acceptor and stimulated by dinitrophenol. The respiratory rate was the same in the Warburg respirometer as that in the polarograph chamber; no effect of agitation was observed. The rates of oxygen consumption over short periods of time were 30–40 μ atoms per hour per mg N for α-keto-glutarate and glutamate, 16 for malate, and 1.3 for β-OH butyrate; except in the last case, satisfactory P/O ratios were obtained. A soluble, heat-stable muscle factor, not precipitated at 100,000 × G, stimulated respiration with each substrate but did not affect phosphorylation. From optical density measurements of muscle mitochondria they appeared to behave as moderately good osmometers; only slight swelling occurred during oxidative phosphorylation, but this increased somewhat when respiration ceased. The osmotic dead space was estimated at 60% of the mitochondrial volume. With polyvinylpyrrolidone the extraparticulate space was found to be 27.8 ± 1.8%.

PIGEON MUSCLE MITOCHONDRIA: ION CONCENTRATIONS, OXIDATIVE PHOSPHORYLATION, SWELLING, AND EXTRAPARTICULATE SPACE

1963 ◽  
Vol 41 (12) ◽  
pp. 2529-2542 ◽  
Author(s):  
G. J. Marcus ◽  
J. F. Manery
Keyword(s):  
Oxygen Consumption ◽  
Oxidative Phosphorylation ◽  
Respiratory Control ◽  
Sodium Potassium ◽  
Density Measurements ◽  
Muscle Mitochondria ◽  
Ion Concentrations ◽  
Heat Stable ◽  
Control Respiration ◽  
Mitochondrial Volume

Metabolically active mitochondria were isolated from pigeon breast muscle in cold 0.45 M sucrose with ATP, EDTA, and tris buffer at pH 7.4. Fresh mitochondria were analyzed for sodium, potassium, nitrogen, iron, and water. They were morphologically intact and showed respiratory control, respiration being limited by the phosphate acceptor and stimulated by dinitrophenol. The respiratory rate was the same in the Warburg respirometer as that in the polarograph chamber; no effect of agitation was observed. The rates of oxygen consumption over short periods of time were 30–40 μ atoms per hour per mg N for α-keto-glutarate and glutamate, 16 for malate, and 1.3 for β-OH butyrate; except in the last case, satisfactory P/O ratios were obtained. A soluble, heat-stable muscle factor, not precipitated at 100,000 × G, stimulated respiration with each substrate but did not affect phosphorylation. From optical density measurements of muscle mitochondria they appeared to behave as moderately good osmometers; only slight swelling occurred during oxidative phosphorylation, but this increased somewhat when respiration ceased. The osmotic dead space was estimated at 60% of the mitochondrial volume. With polyvinylpyrrolidone the extraparticulate space was found to be 27.8 ± 1.8%.

Oxidative phosphorylation in cardiac infarct. Effect of glucose-KCl-insulin solution

1965 ◽  
Vol 209 (2) ◽  
pp. 371-375 ◽  
Author(s):  
Edmundo Calva ◽  
Adela Mujica ◽  
Abdo Bisteni ◽  
Demetrio Sodi-Pallares
Keyword(s):  
Oxygen Consumption ◽  
Oxidative Phosphorylation ◽  
Respiratory Control ◽  
Low Oxygen ◽  
Normal Limits ◽  
Functional Aspects ◽  
Insulin Solution ◽  
Effect Of Glucose ◽  
Infarcted Tissue ◽  
Electrocardiographic Abnormalities

Myocardial infarction was produced in dogs by ligature of the anterior descending coronary artery. Sarcosomes were isolated from normal and infarcted tissue. Oxygen consumption was followed polarographically and adenosine triphosphate was measured as glucose 6-phosphate. One group of animals received a continuous infusion of glucose for 12 hr; another group received "polarizing solution" (glucose-KCl-insulin). Sarcosomes from the first had a low oxygen consumption, no respiratory control, and no oxidative phosphorylation. In contrast, the administration of glucose-KCl-insulin solution maintained practically within normal limits these functional aspects of the sarcosomes. The reversal of electrocardiographic abnormalities by the administration of the polarizing solution coincided with improvement of such biochemical functions. Anesthesia and surgical handling did not appear to modify the behavior of the sarcosomes.

scholarly journals Alteration of Pseudomonas aeruginosa respiration by 4-(1-adamantyl)-phenol derivative

Biologija ◽  
2018 ◽  
Vol 64 (3) ◽  
Author(s):  
Daria M. Dudikova ◽  
Nina O. Vrynchanu ◽  
Valentyna I. Nosar
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Oxygen Consumption ◽  
Oxidative Phosphorylation ◽  
Respiratory Chain ◽  
Structural Integrity ◽  
Respiratory Control ◽  
Endogenous Respiration ◽  
Phenol Derivative ◽  
Succinate Oxidation ◽  
Intact Cells

Derivatives of 4-(1-adamantyl)-phenol are a promising class of antimicrobials affecting the structural integrity and functions of the bacterial cell membrane. The functioning of Pseudomonas aeruginosa respiratory chain and related system of oxidative phosphorylation was investigated before and after treatment with a derivative of 4-(1-adamantyl)-phenol (compound KVM-97). Oxygen consumption was measured polarographically with a Clark-type oxygen electrode. KVM-97 was tested at 0.5× and 1.0× MIC (minimum inhibitory concentration). Specific substrates of the respiratory chain (either 3.0 mM glutamate with 2.0 mM malonate or 3.0 mM succinate with 5.0 μM rotenone) were used. All reactions were stimulated by addition of ADP (0.2 mmol). It was found that at tested concentrations, KVM-97 inhibited the endogenous respiration and substrate oxidation in P. aeruginosa cells. The inhibiting effect was dose-dependent and more pronounced with succinate oxidation rather than glutamate oxidation. The respiratory control index value (RCI) in compound-treated cells was in average 1.5 times lower compared to the intact cells. The decrease in the RCI was related to changing the oxygen uptake rates in state 3 and state 4, which indicate the uncoupling of respiration and oxidative phosphorylation. The data obtained showed that 4-(1-adamantyl)-phenol derivative inhibits oxygen consumption and has uncoupling effects in P. aeruginosa cells.

Sulphide oxidation and oxidative phosphorylation in the mitochondria of the lugworm

1997 ◽  
Vol 200 (1) ◽  
pp. 83-92 ◽  
Author(s):  
S Vökel ◽  
M K Grieshaber
Keyword(s):  
Oxygen Consumption ◽  
Cytochrome C ◽  
Oxidative Phosphorylation ◽  
Cytochrome C Oxidase ◽  
Electron Flow ◽  
Respiratory Control ◽  
Sulphide Oxidation ◽  
Atp Production ◽  
Sulphide Concentration ◽  
Flow Through

Oxygen consumption, ATP production and cytochrome c oxidase activity of isolated mitochondria from body-wall tissue of Arenicola marina were measured as a function of sulphide concentration, and the effect of inhibitors of the respiratory complexes on these processes was determined. Concentrations of sulphide between 6 and 9 µmol l-1 induced oxygen consumption with a respiratory control ratio of 1.7. Production of ATP was stimulated by the addition of sulphide, reaching a maximal value of 67 nmol min-1 mg-1 protein at a sulphide concentration of 8 µmol l-1. Under these conditions, 1 mole of ATP was formed per mole of sulphide consumed. Higher concentrations of sulphide led to a decrease in ATP production until complete inhibition occurred at approximately 50 µmol l-1. The production of ATP with malate and succinate was stimulated by approximately 15 % in the presence of 4 µmol l-1 sulphide, but decreased at sulphide concentrations higher than 15­20 µmol l-1. Cytochrome c oxidase was also inhibited by sulphide, showing half-maximal inhibition at 1.5 µmol l-1 sulphide. Sulphide-induced ATP production was inhibited by antimycin, cyanide and oligomycin but not by rotenone or salicylhydroxamic acid. The present data indicate that sulphide oxidation is coupled to oxidative phosphorylation solely by electron flow through cytochrome c oxidase, whereas the alternative oxidase does not serve as a coupling site. At sulphide concentrations higher than 20 µmol l-1, oxidation of sulphide serves mainly as a detoxification process rather than as a source of energy.

Defective respiration and oxidative phosphorylation in muscle mitochondria of hamsters in the late stages of hereditary muscular dystrophy

1970 ◽  
Vol 48 (9) ◽  
pp. 1037-1042 ◽  
Author(s):  
B. E. Jacobson ◽  
M. C. Blanchaer ◽  
K. Wrogemann
Keyword(s):  
Oxidative Phosphorylation ◽  
Tricarboxylic Acid Cycle ◽  
Respiratory Control ◽  
Test System ◽  
Loose Coupling ◽  
Dystrophic Muscle ◽  
Muscle Mitochondria ◽  
Skeletal Muscle Mitochondria ◽  
Uptake Rates ◽  
Late Stages

Skeletal muscle mitochondria were isolated from 33 dystrophic hamsters of the BIO 14.6 strain, aged 265 ± 13 (S.E.) days, by glass-on-glass homogenization in a sucrose–EDTA medium in the absence of the proteinase Nagarse. These organelles utilized O2 at half the normal rate with pyruvate/fumarate or palmitate as substrate in a manometric test system and exhibited decreased P/O ratios and phosphorylation rates with pyruvate/fumarate. In polarographic experiments the mitochondria from dystrophic muscle, supplemented with L-malate, had significantly depressed O2 uptake rates, respiratory control ratios, and phosphorylation rates with pyruvate, palmityl-L-carnitine, and acetyl-L-carnitine as substrates and low ADP/O ratios with pyruvate and palmityl-L-carnitine. Since the severity of the respiratory depression was similar with the three substrates, it appeared that the defect lay beyond acetyl-CoA in their common degradative pathway. Judging from the rapid rate of succinate and NADH oxidation, the respiratory chain was unimpaired. It was concluded that a defect was present in the tricarboxylic acid cycle of muscle mitochondria isolated without Nagarse from older dystrophic hamsters of the BIO 14.6 strain and that the defect was accompanied by a loose coupling of oxidative phosphorylation.

scholarly journals Effects of sodium and potassium ions on oxidative phosphorylation in relation to respiratory control by a cell-membrane adenosine triphosphatase

1965 ◽  
Vol 97 (2) ◽  
pp. 523-531 ◽  
Author(s):  
DM Blond ◽  
R Whittam
Keyword(s):  
Oxygen Consumption ◽  
Cell Membrane ◽  
Oxidative Phosphorylation ◽  
Adenosine Triphosphatase ◽  
Respiratory Control ◽  
Enzymic Activity ◽  
Cellular Respiration ◽  
Physiological Concentration ◽  
A Cell ◽  
K Concentration

1. A study has been made of the oxygen consumption of kidney homogenates in relation to the ADP concentration as regulated by the cell-membrane adenosine triphosphatase. Stimulation of this enzymic activity by Na(+) and K(+) caused parallel increases in oxygen consumption and ADP concentration. Similarly, inhibition with ouabain caused a parallel fall. The membrane adenosine triphosphatase concerned in active transport therefore appears to regulate respiration through its control of ADP concentration. 2. The respiration of homogenates and mitochondria was also stimulated by K(+) in a way independent of adenosine-triphosphatase activity. It was shown that K(+) facilitates oxidative phosphorylation and the respiratory response to ADP. A K(+) concentration of 25-50mm was needed for maximum oxidative phosphorylation in the presence of physiological concentration of Na(+). Na(+) counteracted K(+) in the effects on mitochondria. It is concluded that K(+) regulates cellular respiration at two structures, one directly in mitochondria, and the second indirectly through control of ADP production at the cell membrane.

Age-related increase in mitochondrial proton leak and decrease in ATP turnover reactions in mouse hepatocytes

1998 ◽  
Vol 275 (2) ◽  
pp. E197-E206 ◽  
Author(s):  
Mary-Ellen Harper ◽  
Shadi Monemdjou ◽  
Jon J. Ramsey ◽  
Richard Weindruch
Keyword(s):  
Oxygen Consumption ◽  
Oxidative Phosphorylation ◽  
Metabolic Control ◽  
Respiratory Control ◽  
Substrate Oxidation ◽  
Membrane Lipid ◽  
Proton Leak ◽  
Control Analysis ◽  
Atp Turnover ◽  
Age Related

Age-related changes in mitochondria, including decreased respiratory control ratios and altered mitochondrial inner membrane lipid composition, led us to study oxidative phosphorylation in hepatocytes from old (30 mo) and young (3 mo) male C57BL/J mice. Top-down metabolic control analysis and its extension, elasticity analysis, were used to identify changes in the control and regulation of the three blocks of reactions constituting the oxidative phosphorylation system: substrate oxidation, mitochondrial proton leak, and the ATP turnover reactions. Resting oxygen consumption of cells from old mice was 15% lower ( P < 0.05) than in young cells. This is explained entirely by a decrease in oxygen consumption supporting ATP turnover reactions. At all values of mitochondrial membrane potential assessed, the proportion of total oxygen consumption used to balance the leak was greater in the old cells than in the young cells. Metabolic control coefficients indicate a shift in control over respiration and phosphorylation away from substrate oxidation toward increased control by leak and by ATP turnover reactions. Control of the actual number of ATP molecules synthesized by mitochondria for each oxygen atom consumed by the ATP turnover and leak reactions was greater in old than in young cells, showing that efficiency in older cells is more sensitive to changes in these two blocks of reactions than in young cells.

scholarly journals Upregulation of COX4-2 via HIF-1α in Mitochondrial COX4-1 Deficiency

Cells ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 452
Author(s):  
Liza Douiev ◽  
Chaya Miller ◽  
Shmuel Ruppo ◽  
Hadar Benyamini ◽  
Bassam Abu-Libdeh ◽  
...  
Keyword(s):  
Oxygen Consumption ◽  
Oxidative Phosphorylation ◽  
Nuclear Localization ◽  
Target Genes ◽  
Compensatory Mechanism ◽  
Terminal Electron Acceptor ◽  
Human Foreskin Fibroblasts ◽  
Mitochondrial Respiratory Complex ◽  
Mrna Transcripts ◽  
Oxphos System

Cytochrome-c-oxidase (COX) subunit 4 (COX4) plays important roles in the function, assembly and regulation of COX (mitochondrial respiratory complex 4), the terminal electron acceptor of the oxidative phosphorylation (OXPHOS) system. The principal COX4 isoform, COX4-1, is expressed in all tissues, whereas COX4-2 is mainly expressed in the lungs, or under hypoxia and other stress conditions. We have previously described a patient with a COX4-1 defect with a relatively mild presentation compared to other primary COX deficiencies, and hypothesized that this could be the result of a compensatory upregulation of COX4-2. To this end, COX4-1 was downregulated by shRNAs in human foreskin fibroblasts (HFF) and compared to the patient’s cells. COX4-1, COX4-2 and HIF-1α were detected by immunocytochemistry. The mRNA transcripts of both COX4 isoforms and HIF-1 target genes were quantified by RT-qPCR. COX activity and OXPHOS function were measured by enzymatic and oxygen consumption assays, respectively. Pathways were analyzed by CEL-Seq2 and by RT-qPCR. We demonstrated elevated COX4-2 levels in the COX4-1-deficient cells, with a concomitant HIF-1α stabilization, nuclear localization and upregulation of the hypoxia and glycolysis pathways. We suggest that COX4-2 and HIF-1α are upregulated also in normoxia as a compensatory mechanism in COX4-1 deficiency.

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