Studies on the availability of intramitochondrial carbamoylphosphate for utilization in extramitochondrial reactions in rat liver |
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Authors: | P J Natale G C Tremblay |
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Institution: | Department of Biochemistry, University of Rhode Island, Kingston, Rhode Island 02881 USA |
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Abstract: | The following observations with isolated mitochondria prepared from rat liver demonstrate that Carbamoylphosphate can readily traverse the mitochondrial membrane: (a) Citrulline synthesis occurs within isolated intact mitochondria at the expense of exogenously added ornithine and 14C]carbamoylphosphate, providing evidence that the initochondrial membrane does not exclude extramitochondrial car bamoylphosphate from penetrating the intramitochondrial matrix, (b) The 14C]carbamoylphosphate synthesized within isolated intact mitochondria from NaH14CO3 by the action of the N-acetyl-l-glutamate-activated carbamoylphosphate synthetase (CPS-I) is equally available for consumption in intramitochondrial and extramitochondrial reactions, as judged by the coupled activity of CPS-I with either intramitochondrial ornithine carbamoyltransferase or extramitochondrial aspartate carbamoyltransferase. The possibility that the coupled action of CPS-I with intramitochondrial ornithine carbamoyltransferase might prevent the export of carbamoylphosphate into the extramitochondrial medium was also examined. The addition of ornithine to the reaction mixture, at concentrations which are optimal for citrulline production, did not reduce carbamoylphosphate export below of the total amount of carbamoylphosphate synthesized. These results indicate that the carbamoylphosphate generated intramitochondrially is not compartment ally excluded from participating in cytoplasmic reactions, and raise the possibility that the intramitochondrial carbamoylphosphate synthetase, CPS-I, may be a significant source of the carbamoylphosphate incorporated into hepatic pyrimidines by the cytoplasmic enzymes of the orotate pathway. |
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Keywords: | To whom requests for reprints should be addressed |
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