Measurement of ATP production and respiratory chain enzyme activities in mitochondria isolated from small muscle biopsy samples.

@article{Wibom2002MeasurementOA,
  title={Measurement of ATP production and respiratory chain enzyme activities in mitochondria isolated from small muscle biopsy samples.},
  author={R. Wibom and L. Hagenfeldt and U. von D{\"o}beln},
  journal={Analytical biochemistry},
  year={2002},
  volume={311 2},
  pages={
          139-51
        }
}
A set of methods suitable for assessment of respiratory chain function in mitochondria isolated from 25mg of muscle is described. This set of methods includes determination of the mitochondrial ATP production rate (MAPR) and the activities of the respiratory chain complexes I, I+III, II+III, and IV and citrate synthase. MAPR is determined with an optimized version of a luminometric method previously described. The optimized method measures 50-220% higher activities than the original method. The… Expand
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References

SHOWING 1-10 OF 34 REFERENCES
An evaluation of the measurement of the activities of complexes I-IV in the respiratory chain of human skeletal muscle mitochondria.
TLDR
Assays which measure complexes I to IV in human skeletal muscle mitochondria are evaluated and optimized to provide sensitive and reliable diagnostic techniques, particularly in situations where a partial interruption at a single complex needs to identified. Expand
A sensitive method for measuring ATP-formation in rat muscle mitochondria.
TLDR
A sensitive method for the measurement of the ATP production rate in isolated skeletal muscle mitochondria is presented, utilizing a reagent based upon firefly luciferase, which emits light proportional to the ATP concentration. Expand
ATP production rate in mitochondria isolated from microsamples of human muscle.
TLDR
Mitochondrial ATP production (MAPR) was determined using a bioluminescence method in mitochondrial preparations of human skeletal muscle using percutaneous muscle biopsy technique and was higher for the combination of substrates than for any individual substrate tested. Expand
Human skeletal muscle mitochondrial capacity.
Under aerobic work, the oxygen consumption and major ATP production occur in the mitochondria and it is therefore a relevant question whether the in vivo rates can be accounted for by mitochondrialExpand
Flux control of cytochrome c oxidase in human skeletal muscle.
TLDR
A tight in vivo control of respiration by COX in human skeletal muscle is indicated, indicating severe rate limitations of oxidative phosphorylation by this enzyme, and may have significant implications for mitochondrial myopathies. Expand
Rate of oxidative phosphorylation in isolated mitochondria from human skeletal muscle: effect of training status.
TLDR
It is concluded that muscle oxidative function can be quantitatively assessed from the respiration of mitochondria isolated from needle biopsy specimens and that QO2 is closely correlated to whole-body VO2 max. Expand
Functional characterization of mitochondrial oxidative phosphorylation in saponin-skinned human muscle fibers.
TLDR
Skinned muscle fibers from M. vastus lateralis with saponin are a suitable object for the replacement of isolated mitochondria in the diagnosis of mitochondrial myopathies and encephalomyopathies. Expand
Human quadricepts muscle mitochondria: A functional characterization
TLDR
Comparison of uncoupled rates of respiration and state 3 rates indicated that the ATP synthesis exerted major flux control over respiration of succinate + glutamate, malate + glutamate and pyruvate + malate, which could be used as a functional assay of ATP synthesis. Expand
Role of mitochondrial calcium transport in the control of substrate oxidation
TLDR
The point of view is that the failure of mitochondrial energy transduction to keep pace with cellular energy needs in some forms of heart failure may involve a failure of [Ca2+]m to be raised adequately to allow the activation of the dehydrogenases, and raises the possibility of therapy based upon partial inhibition of mitochondrial Ca2+ efflux pathways. Expand
Activities of citrate synthase and NAD+-linked and NADP+-linked isocitrate dehydrogenase in muscle from vertebrates and invertebrates.
TLDR
A comparison of the maximal activities of the enzymes with the maximal flux through the cycle suggests that, in insect flight muscle, NAD+-linked isocitrate dehydrogenase catalysed a non-equilibrium reaction and citrate synthease catalyses a near-equ equilibrium reaction. Expand
...
1
2
3
4
...