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The Journal of Neuroscience, April 15, 2000, 20(8):2749-2757
Isolation of Current Components and Partial Reaction Cycles in
the Glial Glutamate Transporter EAAT2
Thomas S.
Otis1 and
Michael P.
Kavanaugh2
1 Department of Neurobiology, University of California,
Los Angeles Medical Center, Los Angeles, California 90095-1763, and
2 Vollum Institute, Portland, Oregon 97202
The kinetic properties of the excitatory amino acid transporter
EAAT2 were studied using rapid applications of L-glutamate to outside-out patches excised from transfected human embryonic kidney
293 cells. In the presence of the highly permeant anion SCN , pulses of glutamate rapidly activated
transient anion channel currents mediated by the transporter. In the
presence of the impermeant anion gluconate, glutamate pulses activated
smaller currents predicted to result from stoichiometric flux of
cotransported ions. Both anion and stoichiometric currents displayed
similar kinetics, suggesting that anion channel gating and
stoichiometric charge movements are linked to early transitions in the
transport cycle. Transporter-mediated anion currents were recorded with
ion and glutamate gradients favoring either unidirectional influx or
exchange. Analysis of deactivation and recovery kinetics in these two
conditions suggests that, after binding, translocation of substrate is
more likely than unbinding under physiological conditions. The kinetic properties of EAAT2, the dominant glutamate transporter in brain astrocytes, distinguish it as an efficient sink for synaptically released glutamate.
Key words:
GLT1; glutamate uptake; anion conductance; EAAT2; astrocyte; glutamate transporter
Copyright © 2000 Society for Neuroscience 0270-6474/00/2082749-09$05.00/0
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