title: Thallium ions can replace both sodium and potassium ions in the glutamate transporter excitatory amino acid carrier 1.
abstract: The excitatory amino acid carrier EAAC1 belongs to a family of glutamate transporters that use the electrochemical transmembrane gradients of sodium and potassium to mediate uphill transport of glutamate into the cell. While the sites of cation interaction with EAAC1 are unknown, two cation binding sites were observed in the crystal structure of the bacterial glutamate transporter homologue GltPh. Although occupied by Tl(+) in the crystal structure, these sites were proposed to be Na(+) binding sites. Therefore, we tested whether Tl(+) has the ability to replace Na(+) also in the mammalian transporters. Our data demonstrate that Tl(+) can bind to EAAC1 with high affinity and mediate a host of different functions. Tl(+) can functionally replace potassium when applied to the cytoplasm and can support glutamate transport current. When applied extracellularly, Tl(+) induces some behavior that mimics that of the Na(+)-bound transporter, such as activation of the cation-induced anion conductance and creation of a substrate binding site, but it cannot replace Na(+) in supporting glutamate transport current. Moreover, our data show a differential effect of mutations to two acidic amino acids potentially involved in cation binding (D367 and D454) on Na(+) and Tl(+) affinity. Overall, our results demonstrate that the ability of the glutamate transporters to interact with Tl(+) is conserved between GltPh and a mammalian member of the transporter family. However, in contrast to GltPh, which does not bind K(+), Tl(+) is more efficient in mimicking K(+) than Na(+) when interacting with the mammalian protein.
authors.affiliation: Department of Chemistry, Binghamton University, 4400 Vestal Parkway East, Binghamton, New York 13850, USA.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Tao
authors.forename: Armanda
authors.initials: A
authors.lastname: Gameiro
authors.forename: Christof
authors.initials: C
authors.lastname: Grewer
mesh_terms: D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002460:Cell Line; D017124:Conserved Sequence; D004553:Electric Conductivity; D050580:Excitatory Amino Acid Transporter 3 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D042541:Intracellular Space / Q000187:drug effects / Q000378:metabolism; D009154:Mutation; D011188:Potassium / Q000378:metabolism*; D051381:Rats; D012964:Sodium / Q000378:metabolism*; D013793:Thallium / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D000596:Amino Acids; D050580:Excitatory Amino Acid Transporter 3; D012964:Sodium; D013793:Thallium; D011188:Potassium