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. 1989 Sep;91(1):272–277. doi: 10.1104/pp.91.1.272

Isolation and Partial Characterization of the Glutamate/Aspartate Transporter from Pea Leaf Mitochondria Using a Specific Monoclonal Antibody 1

Jeevalatha Vivekananda 1, David J Oliver 1
PMCID: PMC1061986  PMID: 16667009

Abstract

A library of monoclonal antibodies directed against the proteins of the inner mitochondrial membrane was screened for antibodies that could bind to the glutamate/aspartate transporter of pea mitochondria and thereby inhibit its activity. One antibody, 2C7, had the property of inhibiting glutamate and aspartate-dependent oxaloacetate metabolism by pea mitochondria without affecting the metabolism of other substrates. The antibody specifically recognized a 21,000 dalton protein, which was tentatively identified as the glutamate/aspartate transporter. The antibody was used to follow the extraction of this protein by Triton X-114 and cardiolipin and the partial purification of the protein by centrifugation and chromatography on hydroxylapatite. The partially purified preparation was reconstituted into azolectin vesicles and shown to catalyze glutamate/glutamate and glutamate/aspartate exchange in an apparently nonelectrogenic manner. The antibody was shown to specifically bind to the glutamate/aspartate exchanger by its ability to inhibit this reconstituted exchange reaction.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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