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Plant Physiology 76:409-413 (1984)
© 1984 American Society of Plant Biologists

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Articles

Characterization of the Transport of Oxaloacetate by Pea Leaf Mitochondria 1

David J. Oliver and Griffin H. Walker

Department of Bacteriology and Biochemistry, University of Idaho, Moscow, Idaho 83843

Mitochondria isolated from pea (Pisum sativum L.) leaves are able to transport the keto acid, oxaloacetate, from the reaction medium into he mitochondrial matrix at high rates. The rate of uptake by the mitochondria was measured as the rate of disappearance of oxaloacetate from the reaction medium as it was reduced by matrix malate dehydrogenase using NADH provided by glycine oxidation. The oxaloacetate transporter was identifed as being distinct from the dicarboxylate and the {alpha}-ketoglutarate transporters because of its inhibitor sensitivities and its inability to interact with other potential substrates. Phthalonate and phthalate were competitive inhibitors of oxaloacetate transport with Ki values of 60 micromolar and 2 millimolar, respectively. Butylmalonate, an inhibitor of the dicarboxylate and {alpha}-ketoglutarate transporters, did not alter the rate of oxaloacetate transport. In addition, a 1000-fold excess of malate, malonate, succinate, {alpha}-ketoglutarate, or phosphate had little effect on the rate of oxaloacetate transport. The Km for the oxaloacetate transporter was about 15 micromolar with a maximum velocity of over 500 nanomoles per milligram mitochondrial protein/min at 25°C. No requirement for a counter ion to move against oxaloacetate was detected and the highest rates of uptake occurred at alkaline pH values. An equivalent transporter has not been reported in animal mitochondria.


1 Funded by a grant from the National Science Foundation (PCM-8204186) and is publication No. 8364 of the Idaho Agricultural Experiment Station.




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Identification of a Novel Transporter for Dicarboxylates and Tricarboxylates in Plant Mitochondria. BACTERIAL EXPRESSION, RECONSTITUTION, FUNCTIONAL CHARACTERIZATION, AND TISSUE DISTRIBUTION
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I. Hanning, K. Baumgarten, K. Schott, and H. W. Heldt
Oxaloacetate Transport into Plant Mitochondria
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