Investigations on the function of creatine kinase in Ehrlich ascites tumor cells.
1. Growth and viability of in vitro cultured Ehrlich ascites tumor cells are not significantly impaired by exogenous creatine up to 40mM. Retardation of cell growth by higher concentrations depends on cell density. 2. Ehrlich cells grown in the presence of high concentrations of creatine accumulate creatine phosphate to high levels (up to 23 nmol/10(6) cells in the presence of 40mM creatine). 3. A nearly complete interruption of glycolytic ATP production or inhibition of the oxidative ATP synthesis reduces the maximal creatine to about 40-50% of controls. 4. Studies on the intracellular distribution of creatine kinase have shown, that the enzyme is only associated with the mitochondrial fraction. Titration of isolated mitochondria with digitonin revealed that the activity is located in the inter-membrane space and partly bound to the outer site of the inner membrane. 5. By growth of Ehrlich cells in creatine-free medium it is possible to obtain "creatine phosphate-depleted" cells (creatine phosphate less than 10% of controls). The growth of creatine phosphate-depleted cells as compared to controls is significantly reduced under energetic stress situations. The protein synthesis of these cells after an energetic stress (lack of glucose and oxygen) is significantly reduced as compared to creatine phosphate containing cells. 6. It is concluded that in these cells creatine kinase/creatine phosphate is a thermodynamic buffer system and not part of an energy shuttle as is postulated for muscle cells.
Physiologisch-Chemisches Institut der Philipps-Universität Marburg.
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S
Becker
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F
Schneider
D000255:Adenosine Triphosphate / Q000032:analysis; D000818:Animals; D002286:Carcinoma, Ehrlich Tumor / Q000201:enzymology*; D003402:Creatine Kinase / Q000502:physiology*; D004072:Digitonin; D005260:Female; D006019:Glycolysis; D007202:Indicators and Reagents; D051379:Mice; D008928:Mitochondria / Q000201:enzymology; D010084:Oxidation-Reduction; D010725:Phosphocreatine / Q000032:analysis; D010766:Phosphorylation; D014407:Tumor Cells, Cultured
D007202:Indicators and Reagents; D010725:Phosphocreatine; D000255:Adenosine Triphosphate; D003402:Creatine Kinase; D004072:Digitonin