rBAT is an amino acid exchanger with variable stoichiometry.
The rBAT protein, when expressed in Xenopus oocytes, was previously shown to reproduce the selectivity of the Na(+)-independent neutral and basic amino acid transport system called bo,+. More recently, the capacity of rBAT to generate a transmembrane current was demonstrated when addition of neutral amino acids stimulated the efflux of cations (presumably basic amino acids) in rBAT-injected oocytes. In the present paper, aminoisobutyric acid (AIB), a neutral amino acid analogue, was shown to induce outward currents (efflux of basic amino acids) through rBAT similar to those caused by alanine in terms of affinity, maximal currents and I-V curves. Despite generating similar currents, the AIB transport rate was more than 30 times lower than that of alanine, thus challenging the assumption that rBAT functions as a classical exchanger. Experiments using a cut-open oocyte voltage clamp demonstrated that AIB was capable of stimulating rBAT-mediated currents from either side of the membrane. AIB, like alanine, was able to stimulate the efflux of radiolabeled alanine and arginine while no rBAT-mediated efflux was measurable in the absence of external rBAT substrates. These results demonstrate that (i) the presence of amino acids is required on both sides of the membrane for rBAT to mediate amino acid flux and thus rBAT must be some type of exchanger but (ii) rBAT-mediated amino acid influx is not stoichiometrically related to the efflux. A model of a "double gated pore" is proposed to account for these properties of rBAT, which contravene standard models of exchangers and other transporters.
Coady
M J
MJ
Département de Physique, Université de Montréal, QC, Canada.
Chen
X Z
XZ
Lapointe
J Y
JY
D000409:Alanine / Q000378:metabolism*; D026923:Amino Acid Transport Systems, Basic; D000596:Amino Acids / Q000378:metabolism; D000621:Aminoisobutyric Acids / Q000378:metabolism*; D000818:Animals; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004553:Electric Conductivity; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D009865:Oocytes / Q000502:physiology; D018408:Patch-Clamp Techniques; D014982:Xenopus laevis
D026923:Amino Acid Transport Systems, Basic; D000596:Amino Acids; D000621:Aminoisobutyric Acids; D002352:Carrier Proteins; D008562:Membrane Glycoproteins; D000409:Alanine