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M202702200v1
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Papers In Press, published online ahead of print April 26, 2002
J. Biol. Chem, 10.1074/jbc.M202702200
Submitted on March 20, 2002
Revised on April 26, 2002
Accepted on April 25, 2002

Identification of a novel transporter for dicarboxylates and tricarboxylates in plant mitochondria: bacterial expression, reconstitution, functional characterization and tissue distribution

Nathalie Picault, Luigi Palmieri, Isabella Pisano, Michael Hodges, and Ferdinando Palmieri

Department of Pharmaco-Biology, University of Bari, Bari 70125

Corresponding Author: fpalm{at}farmbiol.uniba.it

A cDNA from Arabidopsis thaliana and four related cDNAs from Nicotiana tabacum that we have isolated encode hitherto unidentified members of the mitochondrial carrier family. These proteins have been overexpressed in bacteria and reconstitited into phospholipid vesicles. Their transport properties demonstrate that they are orthologs/isoforms of a novel mitochondrial carrier capable of transporting both dicarboxylates (such as malate, oxaloacetate, oxoglutarate and maleate) and tricarboxylates (such as citrate, isocitrate, cis-aconitate and trans-aconitate). The newly identified dicarboxylate-tricarboxylate carrier accepts only the single protonated form of citrate (H-citrate2-) and the unprotonated form of malate (malate2-), and catalyzes obligatory, electroneutral exchanges. Oxoglutarate, citrate and malate are mutually competitive inhibitors, showing Ki close to the respective Km. The carrier is expressed in all plant tissues examined and is largely spread in the plant kingdom. Furthermore, nitrate supply to nitrogen-starved tobacco plants leads to an increase in its mRNA in roots and leaves. The dicarboxylate-tricarboxylate carrier may play a role in important plant metabolic functions requiring organic acid flux to or from the mitochondria, such as N assimilation, export of reducing equivalents from the mitochondria and fatty acid elongation.


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