Kinetic Properties of the Isoenzymes of Human Creatine Phosphokinase

Witteveen, S.A.G.J.; Sobel, Burton E.; DeLuca, Marlene

doi:10.1073/pnas.71.4.1384

Cited by 45 publications

(8 citation statements)

References 17 publications

(10 reference statements)

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“…Because the rate of formation of ATP from creatine phosphate is greater for CK-MB than for CK-MM (45), the acute increase in CK-MB in skeletal muscle after severe exercise may facilitate the intracellular shuttle of energy, thereby enhancing the repletion of local cellular energy stores (8,38). The large increase in plasma CK-MB activity after the marathon race in the present study is similar to those reported previously (3,28,41,43).…”

Section: Resultssupporting

confidence: 91%

Indicators of Cell Breakdown in Plasma of Men During and After a Marathon Race

et al. 1988

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Plasma indicators of muscle cell leakage and of hemolysis were studied in 23 runners before and after a marathon race. Blood samples were drawn from an antecubital vein the morning before the race (baseline), at 3 p.m., i.e., 2 h before the start, on arrival, 12 and 36 h, and 7 days later. Compared with the baseline values, the plasma creatinine phosphokinase MM and MB subfractions, aldolase and glutamicoxaloacetic transaminase activity were increased immediately after the race, rose further 12 h after the marathon, and remained elevated the race, rose further 12 h after the marathon, and remained elevated 36 h and 7 days later. The plasma lactate dehydrogenase activity and myoglobin concentration were increased on arrival and returned to the pre-race activity 7 days after the marathon. Compared with the pre-race values, the plasma haptoglobin concentration was decreased immediately and 12 h after the marathon. Our data show that indicators of muscle cell leakage and of hemolysis in plasma, withdrawn after a marathon race, remained elevated for up to 7 days after the race.

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Section: Resultssupporting

confidence: 91%

Indicators of Cell Breakdown in Plasma of Men During and After a Marathon Race

et al. 1988

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“…Firefly luciferase luminescence has been employed previously for continuous, qualitative measurement of oxidative phosphorylation [16,17], photosynthetic phosphorylation [18,19], and, more recently, a determination of creatine phosphokinase isoenzymes [20]. Such techniques have seen limited application because of the inability to quantitative ATP concentration during the continuous recording.…”

Section: Luminescence and Atp Concentrationmentioning

confidence: 99%

Continuous Measurement and Rapid Kinetics of ATP Synthesis in Rat Liver Mitochondria, Mitoplasts and Inner Membrane Vesicles Determined by Firefly‐Luciferase Luminescence

Lemasters

Hackenbrock

1976

European Journal of Biochemistry

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Oxidative phosphorylation in rat liver mitochondria, the mitoplast or inner membrane-matrix fraction, and inverted inner membrane vesicles was studied with firefly luciferase luminescence. All preparations showed relatively high ATPjO ratios under initial reaction conditions in the presence of Mg2 +, but only intact mitochondria demonstrated significant respiratory control. Mitochondria and mitoplasts, but not vesicles, catalyzed nearly complete conversion of ADP to ATP. Vesicles converted between 50% and 70% of added ADP to ATP establishing a phosphate potential of 9.8 to 10.5 kcal/mol(43.9 kJ/mol). In "loosely coupled" submitochondrial systems which do not display respiratory control, the phosphate potential appears to be a useful parameter of the integrity of the phosphorylating membrane, while ATP/O ratios may be dependent on the relative concentrations of ADP and ATP in the reaction medium.The rapid kinetics of oxidative synthesis of ATP by inverted inner membrane vesicles was investigated in stopped-flow rapid-mixing experiments. After an oxygen pulse to reduced vesicles, ATP synthesis commenced within 20 ms. In antimycin-inhibited vesicles, ATP synthesis resulting from the rapid oxidation of the terminal portion of the respiratory chain commenced within 20 ms and took approximately 100 ms for half completion. Since half oxidation of the terminal portion of the respiratory chain occurs within a few milliseconds, ATP synthesis resulting from such redox reactions must occur over a significantly longer time course than the redox reactions themselves.We have previously described a highly sensitive method using firefly luciferase luminescence to record ATP synthesis quantitatively and continuously in suspensions of mitochondria and submitochondrial particles [I]. Quantitation of ATP using this method is based upon a theoretical and experimental analysis of the kinetics of luciferase luminescence. In this communication, we describe the use of the luminescence method to characterize the oxidative synthesis of ATP in three mitochondria1 and submitochondrial systems : isolated rat liver mitochondria ; the mitoplast or inner membrane-matrix fraction of rat h e r mitochondria ; and sonicated inner membrane vesicles prepared from the purified mitoplast fraction.Our objectives are several-fold. One is to establish further the accuracy and reliability of the luminescence method for the continuous measurement of ATP in mitochondrial and submitochondrial suspensions during oxidative phosphorylation. Another is to examine various indices of oxidative phosphorylation such as acceptor control, ATPjO ratio and phosphate potential with respect to their relative sensitivity to the functional and structural integrity of the phosphorylating membrane. A further important objective is to study the rapid kinetics of the oxidative synthesis of ATP. EXPERIMENTAL PROCEDUREPurified firefly luciferase and synthetic firefly luciferin were obtained from DuPont Corp. Rat liver mitochondria [2,3] were fractionated to the mitoplast or inner m...

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“…The CK activity was measured with a luminescence technique based on the method proposed by Witteveen et al ( 1974) and Lundin (1 978). The prisms from Torpedo electric organ were homogenized in 100 m M Tris, 1 mM EDTA, 5 mM Mg acetate, 2 M mercaptoethanol, pH 8.…”

Section: Activity Of Ckmentioning

confidence: 99%

Energy Metabolism and Quantal Acetylcholine Release: Effects of Botulinum Toxin, l‐Fluoro‐2,4‐Dinitrobenzene, and Diamide in the Torpedo Electric Organ

Dunant¹,

Loctin²,

Marsal³

et al. 1988

Journal of Neurochemistry

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In the Torpedo electric organ, a modified nerve-muscle system, type A botulinum toxin blocked the release of acetylcholine (ACh) quanta, both neurally evoked and spontaneous. At the same time, the toxin increased the release of a class of small miniature potentials (the subminiature potentials), reduced the ATP and more the creatine phosphate content of the tissue, and impaired the activity of creatine kinase (CK). Thus, we compared this pattern of changes with those provoked by 1-fluoro-2,4-dinitrobenzene (FDNB), an efficient inhibitor of CK. As expected, FDNB rapidly inactivated CK, which resulted in a profound depletion of ATP whereas the stores of creatine phosphate were preserved. In addition, FDNB caused conspicuous morphological alterations of nerve endings and ACh depletion. This agent also suppressed evoked and spontaneous quantal release whereas the occurrence of subminature potentials was markedly increased. Diamide, a penetrating thiol oxidizing substance, provoked first a transient rise in quantal ACh release and then blockade of transmission with, again, production of a large number of subminiature potentials. Creatine phosphate was depleted in the tissue by diamide, the ATP content reduced, and CK activity partly inhibited. The morphology of nerve terminals did not show obvious changes with either diamide or botulinum toxin at the stage of transmission failure. Although the three poisons acted by different mechanisms, this resulted in a rather similar pattern of physiological changes: failure of quantal release and enhancement of subquantal release. These results and experiments on synaptosomes indicated that CK inhibition was probably a crucial mechanism for FDNB but not for diamide or botulinum intoxication.(ABSTRACT TRUNCATED AT 250 WORDS)

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Kinetic Properties of the Isoenzymes of Human Creatine Phosphokinase

Cited by 45 publications

References 17 publications

Indicators of Cell Breakdown in Plasma of Men During and After a Marathon Race

Indicators of Cell Breakdown in Plasma of Men During and After a Marathon Race

Continuous Measurement and Rapid Kinetics of ATP Synthesis in Rat Liver Mitochondria, Mitoplasts and Inner Membrane Vesicles Determined by Firefly‐Luciferase Luminescence

Energy Metabolism and Quantal Acetylcholine Release: Effects of Botulinum Toxin, l‐Fluoro‐2,4‐Dinitrobenzene, and Diamide in the Torpedo Electric Organ

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