title: Identification of a novel Na+-independent acidic amino acid transporter with structural similarity to the member of a heterodimeric amino acid transporter family associated with unknown heavy chains.
abstract: We identified a novel Na(+)-independent acidic amino acid transporter designated AGT1 (aspartate/glutamate transporter 1). AGT1 exhibits the highest sequence similarity (48% identity) to the Na(+)-independent small neutral amino acid transporter Asc (asc-type amino acid transporter)-2 a member of the heterodimeric amino acid transporter family presumed to be associated with unknown heavy chains (Chairoungdua, A., Kanai, Y., Matsuo, H., Inatomi, J., Kim, D. K., and Endou, H. (2001) J. Biol. Chem. 276, 49390-49399). The cysteine residue responsible for the disulfide bond formation between transporters (light chains) and heavy chain subunits of the heterodimeric amino acid transporter family is conserved for AGT1. Because AGT1 solely expressed or coexpressed with already known heavy chain 4F2hc (4F2 heavy chain) or rBAT (related to b(0,+)-amino acid transporter) did not induce functional activity, we generated fusion proteins in which AGT1 was connected with 4F2hc or rBAT. The fusion proteins were sorted to the plasma membrane and expressed the Na(+)-independent transport activity for acidic amino acids. Distinct from the Na(+)-independent cystine/glutamate transporter xCT structurally related to AGT1, AGT1 did not accept cystine, homocysteate, and l-alpha-aminoadipate and exhibited high affinity to aspartate as well as glutamate, suggesting that the negative charge recognition site in the side chain-binding site of AGT1 would be closer to the alpha-carbon binding site compared with that of xCT. The AGT1 message was predominantly expressed in kidney. In mouse kidney, AGT1 protein was present in the basolateral membrane of the proximal straight tubules and distal convoluted tubules. In the Western blot analysis, AGT1 was detected as a high molecular mass band in the nonreducing condition, whereas the band shifted to a 40-kDa band corresponding to the AGT1 monomer in the reducing condition, suggesting the association of AGT1 with other protein via a disulfide bond. The finding of AGT1 and Asc-2 has established a new subgroup of the heterodimeric amino acid transporter family whose members associate not with 4F2hc or rBAT but with other unknown heavy chains.
authors.affiliation: Department of Pharmacology and Toxicology, Kyorin University School of Medicine, 6-20-2 Shinkawa, Mitaka, Tokyo 181-8611, Japan.
authors.forename: Hirotaka
authors.initials: H
authors.lastname: Matsuo
authors.forename: Yoshikatsu
authors.initials: Y
authors.lastname: Kanai
authors.forename: Ju Young
authors.initials: JY
authors.lastname: Kim
authors.forename: Arthit
authors.initials: A
authors.lastname: Chairoungdua
authors.forename: Do Kyung
authors.initials: DK
authors.lastname: Kim
authors.forename: Jun
authors.initials: J
authors.lastname: Inatomi
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Shigeta
authors.forename: Hisako
authors.initials: H
authors.lastname: Ishimine
authors.forename: Sophapun
authors.initials: S
authors.lastname: Chaekuntode
authors.forename: Kittipong
authors.initials: K
authors.lastname: Tachampa
authors.forename: Hye Won
authors.initials: HW
authors.lastname: Choi
authors.forename: Ellappan
authors.initials: E
authors.lastname: Babu
authors.forename: Jun
authors.initials: J
authors.lastname: Fukuda
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Endou
mesh_terms: D000595:Amino Acid Sequence; D026905:Amino Acid Transport Systems / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D017931:DNA Primers; D007668:Kidney / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid; D012964:Sodium / Q000378:metabolism*; D014981:Xenopus
chemical_list: D026905:Amino Acid Transport Systems; D017931:DNA Primers; C486757:SLC7A13 protein, human; C459184:Slc7a13 protein, mouse; D012964:Sodium