Cloning and Functional Expression of a Human Na super(+) and Cl super(-)-dependent Neutral and Cationic Amino Acid Transporter B super(0+)

A Na super(+)-dependent neutral and cationic amino acid transport system (B super(0+)) plays an important role in many cells and tissues; however, the molecular basis for this transport system is still unknown. To identify new transporters, the expressed sequence tag database was queried, and cDNA fragments with sequence similarity to the Na super(+)/Cl super(-)-dependent neurotransmitter transporter family were identified. Based on these sequences, rapid amplification of cDNA ends of human mammary gland cDNA was used to obtain a cDNA of 4.5 kilobases (kb). The open reading frame encodes a 642-amino acid protein named amino acid transporter B super(0+). Human ATB super(0+) (hATB super(0+)) is a novel member of the Na super(+)/Cl super(-)-dependent neurotransmitter transporter family with the highest sequence similarity to the glycine and proline transporters. Northern blot analysis identified transcripts of ~4.5 kb and ~2 kb in the lung. Another tissue survey suggests expression in the trachea, salivary gland, mammary gland, stomach, and pituitary gland. Electrophysiology and radiolabeled amino acid uptake measurements were used to functionally characterize the transporter expressed in Xenopus oocytes. hATB super(0+) was found to transport both neutral and cationic amino acids, with the highest affinity for hydrophobic amino acids and the lowest affinity for proline. Amino acid transport was Na super(+) and Cl super(-)-dependent and was attenuated in the presence of 2-aminobicyclo-[2.2.1]-heptane-2- carboxylic acid a system B super(0+) inhibitor. These characteristics are consistent with system B super(0+) amino acid transport. Thus hATB super(0+) is the first cloned B super(0+) amino acid transporter.