scholarly journals Effect of protonmotive force on the relative proton stoichiometries of the mitochondrial proton pumps

1991 ◽  
Vol 275 (1) ◽  
pp. 75-80 ◽  
Author(s):  
R P Hafner ◽  
M D Brand
Keyword(s):  
Oxygen Consumption ◽  
Oxygen Atom ◽  
Respiration Rate ◽  
Proton Leak ◽  
Protonmotive Force ◽  
Proton Pumps ◽  
Isolated Mitochondria

The rate of phosphorylation of ADP by isolated mitochondria respiring on succinate was set by addition of ATP, ADP or ADP plus malonate. We measured the rates of phosphorylation and respiration and the protonmotive force under each of these conditions. We measured the oxygen consumption required to drive the proton leak at the protonmotive force reached under each condition and subtracted it from the respiration rate during phosphorylation to determine the oxygen consumption driving phosphorylation. By dividing the rate of phosphorylation by the rate of respiration driving phosphorylation we calculated the mechanistic P/O ratio (number of molecules of ADP phosphorylated per oxygen atom reduced). This ratio was the same at high, intermediate and low values of protonmotive force, indicating that the relative stoichiometries of the mitochondrial protonmotive-force-producing and protonmotive-force-consuming pumps (i.e. H+/O:H+/ATP) are independent of the protonmotive force. This greatly weakens the case for a decrease in stoichiometry, or ‘slip’, in the mitochondrial proton pumps at high protonmotive force.

scholarly journals Experimental discrimination between proton leak and redox slip during mitochondrial electron transport

1994 ◽  
Vol 297 (1) ◽  
pp. 27-29 ◽  
Author(s):  
M D Brand ◽  
L F Chien ◽  
P Diolez
Keyword(s):  
Oxygen Consumption ◽  
Proton Leak ◽  
Protonmotive Force ◽  
Proton Pumps ◽  
Proton Conductance ◽  
Turnover Rates ◽  
High Turnover ◽  
Mitochondrial Inner Membrane ◽  
Rate Of Oxygen Consumption ◽  
The Relationship

By measuring the relationship between protonmotive force and the increment in oxygen consumption by mitochondria treated with submaximal amounts of uncoupler, we have experimentally tested four different models of imperfect coupling of oxidative phosphorylation. The results show that the increased rate of oxygen consumption at high protonmotive force is explained entirely by the dependence on protonmotive force of the passive proton leak conductance of the mitochondrial inner membrane. There is no measurable contribution from redox-slip reactions in the proton pumps caused by high protonmotive force. Neither is there any contribution from increased proton conductance of the membrane or increased redox slip in the respiratory chain caused by high turnover rates of the complexes.

scholarly journals Control of the effective P/O ratio of oxidative phosphorylation in liver mitochondria and hepatocytes

1993 ◽  
Vol 291 (3) ◽  
pp. 739-748 ◽  
Author(s):  
M D Brand ◽  
M E Harper ◽  
H C Taylor
Keyword(s):  
Oxygen Consumption ◽  
Liver Mitochondria ◽  
Substrate Oxidation ◽  
Proton Leak ◽  
Rat Liver Mitochondria ◽  
Protonmotive Force ◽  
Control Analysis ◽  
Control Coefficient ◽  
Oxidation Reactions ◽  
In Cells

The control exerted by substrate oxidation reactions, by ATP turnover and by the proton leak over the oxygen consumption rate, the phosphorylation rate, the proton leak rate and the protonmotive force (delta p) in isolated rat liver mitochondria under a range of conditions between non-phosphorylating (State 4) and maximum phosphorylation (State 3) was investigated by using the top-down approach of metabolic control analysis. The experiments were carried out with saturating concentrations of the substrates succinate, glutamate with malate, or pyruvate with malate. The distribution of control was very similar with each of the three substrates. The effective P/O ratio (i.e. not corrected for leak reactions) was also measured; it varied from zero in State 4 to 80-90% of the maximum theoretical P/O ratio in State 3. Under most conditions control over the effective P/O ratio was shared between proton leak (which had negative control) and the phosphorylating subsystem (which had roughly equal positive control); near State 4, substrate oxidation reactions also acquired some control over this ratio. In resting hepatocytes the effective P/O ratio was only 50% of its maximum theoretical value, corresponding to an effective P/O ratio of only 1.3 for complete oxidation of glucose. The effective P/O ratio for intracellular mitochondrial oxygen consumption was 64% of the maximum value. The control coefficient of the mitochondrial proton leak over the effective P/O ratio in cells was -0.34; the control coefficient of phosphorylation reactions over this ratio was 0.31 and the control coefficient of substrate oxidation reactions over the ratio was 0.03, showing how the coupling efficiency in cells can respond sensitively to agents that change the proton leak or the ATP demand, but not to those that change substrate oxidation.

ConA induced changes in energy metabolism of rat thymocytes

1992 ◽  
Vol 12 (5) ◽  
pp. 381-386 ◽  
Author(s):  
F. Buttgereit ◽  
M. D. Brand ◽  
M. Müller
Keyword(s):  
Protein Synthesis ◽  
Oxygen Consumption ◽  
Energy Metabolism ◽  
Dna Synthesis ◽  
Atp Synthesis ◽  
Proton Leak ◽  
Ehrlich Ascites ◽  
Activated State ◽  
Ehrlich Ascites Tumour ◽  
Induced Changes

The influence of ConA on the energy metabolism of quiescent rat thymocytes was investigated by measuring the effects of inhibitors of protein synthesis, proteolysis, RNA/DNA synthesis, Na+K+-ATPase, Ca2+-ATPase and mitochondrial ATP synthesis on respiration. Only about 50% of the coupled oxygen consumption of quiescent thymocytes could be assigned to specific processes using two different media. Under these conditions the oxygen is mainly used to drive mitochondrial proton leak and to provide ATP for protein synthesis and cation transport, whereas oxygen consumption to provide ATP for RNA/DNA synthesis and ATP-dependent proteolysis was not measurable. The mitogen ConA produced a persistent increase in oxygen consumption by about 30% within seconds. After stimulation more than 80% of respiration could be assigned to specific processes. The major oxygen consuming processes of ConA-stimulated thymocytes are mitochondrial proton leak, protein synthesis and Na+K+-ATPase with about 20% each of total oxygen consumption, while Ca2+-ATPase and RNA/DNA synthesis contribute about 10% each. Quiescent thymocytes resemble resting hepatocytes in that most of the oxygen consumption remains unexplained. In constrast, the pattern of energy metabolism in stimulated thymocytes is similar to that described for Ehrlich Ascites tumour cells and splenocytes, which may also be in an activated state. Most of the oxygen consumption is accounted for, so the unexplained process(es) in unstimulated cells shut(s) off on stimulation.

Effect of growth rate on the physiological rates of a chemostat-grown rotifer Encentrum linnhei

1988 ◽  
Vol 68 (1) ◽  
pp. 165-177 ◽  
Author(s):  
J. M. Scott
Keyword(s):  
Growth Rate ◽  
Oxygen Consumption ◽  
Steady State ◽  
Respiration Rate ◽  
Clearance Rate ◽  
Ingestion Rate ◽  
Physiological State ◽  
Growth Efficiency ◽  
Theoretical Figure ◽  
Marine Rotifer

The physiological rates of a normally omnivorous marine rotifer, Encentrum linnhei, were measured under the steady-state chemostat conditions in which the physiological state of the food-algae was kept constant whilst the rotifer growth rate was changed to preset levels. The specific clearance rate ranged between 50 and 100 μl/μg rotifer C/day (1.5–3.0 μ/rot/day) and varied hyperbolically with growth rate, a similar curve was obtained with the specific ingestion rate which varied between 1–2 μg C/μg rot C/day. A mean respiration rate of 0.45 μg C/μg rot C/day was obtained from oxygen consumption measurements. About 60‰ of ingested energy was found to be egested as paniculate matter and 9–4 °0 dissipated as heat, the latter comparing with a theoretical figure of 4–5‰.From rates, transfer efficiencies were obtained giving a mean net growth efficiency (K2) of 38‰ and a mean overall growth efficiency (K1 of 15‰. A curvilinear increase of Kl with growth rate contrasts with linear and hyperbolic responses found with brachionid rotifers.

scholarly journals The Effect of Ionic Copper on the Oxygen Consumption of Gammarus Pulex and Polycelis Nigra

1941 ◽  
Vol 18 (2) ◽  
pp. 153-161
Author(s):  
J. R. ERICHSEN JONES
Keyword(s):  
Oxygen Consumption ◽  
Respiration Rate ◽  
Copper Sulphate ◽  
Muscular Activity ◽  
Gammarus Pulex ◽  
Cellular Respiration ◽  
Appreciable Effect ◽  
Normal Value ◽  
Ionic Copper ◽  
Effect Of Copper

The suggestion has been put forward that the oligodynamic action of certain heavy metals is the result of the destruction or inactivation of substances essential for cellular respiration. In a study of the effect of copper sulphate solutions on the oxygen consumption of Polycelis nigra it is found that solutions of concentration 0.001-0.01 N, fatal in 2 hr. or less, induce a marked preliminary rise in the respiration rate; this appears to be due to the inhibition of ciliary locomation and increased muscular activity. A similar increase is produced by increasing the activity of the animals by mechanical means, or by a muscle stimulant (barium chloride). Over the latter part of the survival time the respiration rate drops rapidly and disintegration of the animals begins when it falls to about 60% of the normal value. A 0.0004 N copper sulphate solutions does not inhibit ciliary locomotion, does not stimulate muscular activity, and the oxygen consumption undergoes a stedy decline. A 0.0002 N NaCN solution rapidly depresses the respiration rate to less than 20% of the normal value, but is not fatal, the animals surviving over 4 days. Hydrogen ions, at the concentrations resulting from the hydrolysis of the salt, have no appreciable effect on the oxygen consumption, but at lethal concentrations (pH 2.6, 2.8) effect a speedy depression. The results suggest that the depression of respiration rate observed is insufficient to account for the death of the animals, and is no more than a symptom of the toxic process. A similar general result was obtained in experiments with silver nitrate and mercuric chloride, and also in experiments on the comparative effect of copper sulphate and sodium cyanide solutions on the oxygen consumption of Gammarus pulex.

ConA induced changes in energy metabolism of rat thymocytes

1992 ◽  
Vol 12 (2) ◽  
pp. 109-114 ◽  
Author(s):  
F. Buttgereit ◽  
M. D. Brand ◽  
M. Müller
Keyword(s):  
Protein Synthesis ◽  
Oxygen Consumption ◽  
Energy Metabolism ◽  
Dna Synthesis ◽  
Atp Synthesis ◽  
Proton Leak ◽  
Ehrlich Ascites ◽  
Activated State ◽  
Ehrlich Ascites Tumour ◽  
Induced Changes

The influence of ConA on the energy metabolism of quiescent rat thymocytes was investigated by measuring the effects of inhibitors of protein synthesis, proteolysis, RNA/DNA synthesis, Na+K+-ATPase, Ca2+-ATPase and mitochondrial ATP synthesis on respiration. Only about 50% of the coupled oxygen consumption of quiescent thymocytes could be assigned to specific processes using two different media. Under these conditions the oxygen is mainly used to drive mitochondrial proton leak and to provide ATP for protein synthesis and cation transport, whereas oxygen consumption to provide ATP for RNA/DNA synthesis and ATP-dependent proteolysis was not measurable. The mitogen ConA produced a persistent increase in oxygen consumption by about 30% within seconds. After stimulation more than 80% of respiration could be assigned to specific processes. The major oxygen consuming processes of ConA-stimulated thymocytes are mitochondrial proton leak, protein synthesis and Na+K+-ATPase with about 20% each of total oxygen consumption, while Ca2+-ATPase and RNA/DNA synthesis contribute about 10% each. Quiescent thymocytes resemble resting hepatocytes in that most of the oxygen consumption remains unexplained. In contrast, the pattern of energy metabolism in stimulated thymocytes is similar to that described for Ehrlich Ascites tumour cells and splenocytes, which may also be in an activated state. Most of the oxygen consumption is accounted for, so the unexplained process(es) in unstimulated cells shut(s) off on stimulation.

Oligomycin sensitivity of mitochondrial F1F0-ATPase in diabetes-prone BHE/Cdb rats

1999 ◽  
Vol 277 (4) ◽  
pp. E702-E707 ◽  
Author(s):  
Sook-Bae Kim ◽  
Carolyn D. Berdanier
Keyword(s):  
Mitochondrial Dna ◽  
Oxygen Consumption ◽  
Atpase Activity ◽  
Dna Sequence ◽  
Response Curve ◽  
Human Fibroblasts ◽  
Sprague Dawley ◽  
Isolated Mitochondria ◽  
Sprague Dawley Rats ◽  
Atpase 6

BHE/Cdb and Sprague-Dawley rats differ in their mitochondrial DNA sequence for the ATPase 6 (“subunit a”) gene. Base substitutions in this sequence result in the substitution of asparagine for aspartate at position 101 and the substitution of serine for leucine at position 129. Differences in sensitivity to oligomycin were observed. When the isolated F1F0-ATPase complex was studied and ATPase activity was assessed, that which was isolated from the BHE/Cdb rats was less sensitive to oligomycin inhibition than that which was isolated from the Sprague-Dawley rats. In contrast, when oxygen consumption was measured [oxygen phosphorylation (OXPHOS)] and a dose-response curve was generated with isolated mitochondria from these two strains, there was a shift to the left for the BHE/Cdb rat mitochondria. These mitochondria were more sensitive to oligomycin inhibition of OXPHOS than were mitochondria isolated from Sprague-Dawley rats. The OXPHOS results are consistent with those from human fibroblasts having either a normal or mutated ATPase 6 gene.

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