Opposite directed currents induced by the transport of dibasic and neutral amino acids in Xenopus oocytes expressing the protein rBAT.
Voltage- and current-clamp studies have been performed on a renal and intestinal protein (rBAT) which induces transport for neutral and dibasic amino acids when expressed in Xenopus oocytes. In current-clamp mode, superfusion with L-leucine caused a hyperpolarization while superfusion with L-arginine depolarized the oocyte. Accordingly, in voltage-clamp experiments dibasic amino acids and neutral amino acids induced inward and outward currents, respectively. The relationship between currents and substrate concentrations could be fitted by simple Michaelis-Menten kinetics. Currents induced by L-arginine and L-leucine were also voltage-dependent. pH changes from 6.25 to 8.75 did not affect the currents induced by saturating concentrations of L-arginine and L-leucine, but reversed the direction of L-histidine-induced currents from inward to outward. The reversal potentials as well as the apparent Km for L-histidine-induced currents were altered by the ambient pH. Currents induced by individual amino acids decreased during extended superfusion periods. However, extended superfusion with neutral amino acids increased dibasic amino acid induced currents, while prior superfusion with dibasic amino acid resulted in an increase of currents induced by neutral amino acids. The reversal potentials for L-leucine- and L-arginine-induced currents were depending on their intra- (after preloading) and extracellular concentrations. In conclusion, rBAT-mediated transport of neutral and dibasic amino acids is associated with net outward or inward currents, respectively, which may be caused by an exchange of neutral with dibasic amino acids.
Busch
A E
AE
Institute of Physiology, Eberhard-Karls-Universität Tübingen, Federal Republic of Germany.
Herzer
T
T
Waldegger
S
S
Schmidt
F
F
Palacin
M
M
Biber
J
J
Markovich
D
D
Murer
H
H
Lang
F
F
D026923:Amino Acid Transport Systems, Basic; D000818:Animals; D001120:Arginine / Q000378:metabolism*; D001692:Biological Transport; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004553:Electric Conductivity; D006639:Histidine / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007930:Leucine / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D008564:Membrane Potentials / Q000187:drug effects; D008839:Microelectrodes; D009865:Oocytes; D011994:Recombinant Proteins / Q000378:metabolism; D014982:Xenopus laevis
D026923:Amino Acid Transport Systems, Basic; D002352:Carrier Proteins; D008562:Membrane Glycoproteins; D011994:Recombinant Proteins; D006639:Histidine; D001120:Arginine; D007930:Leucine