title: Regulation of the glutamine transporter SN1 by extracellular pH and intracellular sodium ions.
abstract: The glutamine transporter SN1 has recently been identified as one of the major glutamine transporters in hepatocytes and brain astrocytes. It appears to be the molecular correlate of system N amino acid transport. Two different transport mechanisms have been proposed for this transporter. These are an electroneutral mechanism, in which glutamine uptake is coupled to an exchange of 1Na+ and 1H+, or an electrogenic mechanism coupled to the exchange of 2Na+ against 1H+. This study was performed to solve these discrepancies and to investigate the reversibility of the transporter. When SN1 was expressed in Xenopus laevis oocytes, glutamine uptake was accompanied by a cotransport of 2-3 Na+ ions as determined by 22Na+ fluxes. However, at the same time a rapid release of intracellular Na+ was observed indicating an active exchange of Na+ ions. The driving force of the proton electrochemical gradient was equivalent to that of the sodium electrochemical gradient. Acidification of the extracellular medium caused the transporter to run in reverse and to release glutamine. Determination of accumulation ratios at different driving forces were in agreement with an electroneutral 1Na+-glutamine cotransport-1H+ antiport. Inward currents that were observed during glutamine uptake were much smaller than expected for a stoichiometric cotransport of charges. A slippage mode in the transporter mechanism and pH-regulated endogenous oocyte cation channels are likely to contribute to the observed currents.
authors.affiliation: School of Biochemistry & Molecular Biology, Australian National University, Canberra ACT 0200, Australia.
authors.forename: Angelika
authors.initials: A
authors.lastname: Bröer
authors.forename: Alexandra
authors.initials: A
authors.lastname: Albers
authors.forename: Iwan
authors.initials: I
authors.lastname: Setiawan
authors.forename: Robert H
authors.initials: RH
authors.lastname: Edwards
authors.forename: Farrukh A
authors.initials: FA
authors.lastname: Chaudhry
authors.forename: Florian
authors.initials: F
authors.lastname: Lang
authors.forename: Carsten A
authors.initials: CA
authors.lastname: Wagner
authors.forename: Stefan
authors.initials: S
authors.lastname: Bröer
mesh_terms: D026921:Amino Acid Transport Systems, Neutral; D000818:Animals; D001692:Biological Transport / Q000502:physiology; D055598:Chemical Phenomena; D002621:Chemistry; D004563:Electrochemistry; D005110:Extracellular Space / Q000378:metabolism*; D005260:Female; D005973:Glutamine / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007425:Intracellular Membranes / Q000378:metabolism*; D007477:Ions; D008433:Mathematics; D026901:Membrane Transport Proteins / Q000378:metabolism*; D009865:Oocytes; D011522:Protons; D012964:Sodium / Q000378:metabolism / Q000502:physiology*; D014982:Xenopus laevis
chemical_list: D026921:Amino Acid Transport Systems, Neutral; D007477:Ions; D026901:Membrane Transport Proteins; D011522:Protons; C403222:system N protein 1; D005973:Glutamine; D012964:Sodium