Ammonium ions enhance the flow through the pyruvate dehydrogenase in Ehrlich ascites tumor cells |
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Authors: | J M Carrascosa P Martinez I N de Castro |
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Affiliation: | Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias, Universidad Autonoma, Madrid (34), Spain |
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Abstract: | The flow through pyruvate dehydrogenase was assayed in glycolysing cells by the evolution of 14CO2 from [1-14C] pyruvate. Parallel incubations were carried out in high bicarbonate buffer (25 mM) and in bicarbonate-free buffer. The activation of the complex by NH+4 was only observed in high bicarbonate buffer, because the dilution of labelled CO2 in the presence of an excess of bicarbonate enables the quantitative determination of labelled CO2 evolved from pyruvate in the decarboxylase step. In the bicarbonate-free buffer the activation of the complex was not observed, because the 14CO2 evolved from pyruvate was consumed by biosynthetic processes inside the cell. On the contrary in isolated hepatocytes the NH+4 activation of the pyruvate dehydrogenase was observed in both buffers. In Ehrlich ascites cells, in common with other mammalian tissues, pyruvate dehydrogenase activity was found to be inversely correlated to the intramitochondrial ATP/ADP ratio. |
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Keywords: | ammonium pyruvate déshydrogénase cellules tumorales d'ascites cellules hépatiques isolées ammonium pyruvate dehydrogenase ascites tumor cells isolated hepatocytes |
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