Molecular cloning of Aralar, a new member of the mitochondrial carrier superfamily that binds calcium and is present in human muscle and brain.
We have identified a new calcium-dependent subfamily of mitochondrial carrier proteins with members in Saccharomyces cerevisiae, Caenorhabditis elegans, and various mammalian species. The members of this subfamily have a bipartite structure: a carboxyl-terminal half with the characteristic features of the mitochondrial solute carrier superfamily and an amino-terminal extension harboring various EF-hand domains. A member of this subfamily (that we have termed Aralar) was cloned from a human heart cDNA library. The corresponding cDNA comprises an open reading frame of 2037 base pairs encoding a polypeptide of 678 amino acids. The carboxyl-terminal half of Aralar (amino acids 321-678) has high similarity with the oxoglutarate, citrate, and adenine nucleotide carriers (28-29% identity), whereas the amino-terminal half (amino acids 1-320) contains three canonical EF-hands. Aralar amino-terminal half was shown to bind calcium by 45Ca2+ overlay and calcium-dependent mobility shift assays. The subcellular localization of the protein in COS cells transfected with Aralar was exclusively mitochondrial. Antibodies against Aralar amino-terminal fusion protein recognized a 70-kDa protein in brain mitochondrial fractions. Northern blot analysis showed that the protein was expressed in heart, brain, and skeletal muscle. The domain structure, mitochondrial localization, and presence in excitable tissues suggests a possible function of Aralar as calcium-dependent mitochondrial solute carrier.
Departamento de Biologia Molecular, Centro de Biologia Molecular Severo Ochoa, Consejo Superior de Investigaciones Científicas, Universidad Autónoma de Madrid, 28049-Madrid, Spain.
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del Arco
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Satrústegui
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001921:Brain; D019556:COS Cells; D002118:Calcium / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D002451:Cell Compartmentation; D018076:DNA, Complementary / Q000235:genetics; D006801:Humans; D026901:Membrane Transport Proteins; D008928:Mitochondria / Q000378:metabolism*; D008931:Mitochondria, Muscle; D033681:Mitochondrial Membrane Transport Proteins; D024101:Mitochondrial Proteins; D008969:Molecular Sequence Data; D005810:Multigene Family; D018482:Muscle, Skeletal; D009206:Myocardium; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D014018:Tissue Distribution
D002135:Calcium-Binding Proteins; D018076:DNA, Complementary; D026901:Membrane Transport Proteins; D033681:Mitochondrial Membrane Transport Proteins; D024101:Mitochondrial Proteins; C114606:SLC25A12 protein, human; D002118:Calcium