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Transport direction determines the kinetics of substrate transport by the glutamate transporter EAAC1.

Abstract：

:Glutamate transport by the excitatory amino acid carrier EAAC1 is known to be reversible. Thus, glutamate can either be taken up into cells, or it can be released from cells through reverse transport, depending on the electrochemical gradient of the co- and countertransported ions. However, it is unknown how fast and by which reverse transport mechanism glutamate can be released from cells. Here, we determined the steady- and pre-steady-state kinetics of reverse glutamate transport with submillisecond time resolution. First, our results suggest that glutamate and Na(+) dissociate from their cytoplasmic binding sites sequentially, with glutamate dissociating first, followed by the three cotransported Na(+) ions. Second, the kinetics of glutamate transport depend strongly on transport direction, with reverse transport being faster but less voltage-dependent than forward transport. Third, electrogenicity is distributed over several reverse transport steps, including intracellular Na(+) binding, reverse translocation, and reverse relocation of the K(+)-bound EAAC1. We propose a kinetic model, which is based on a "first-in-first-out" mechanism, suggesting that glutamate association, with its extracellular binding site as well as dissociation from its intracellular binding site, precedes association and dissociation of at least one Na(+) ion. Our model can be used to predict rates of glutamate release from neurons under physiological and pathophysiological conditions.

journal_name

Proc Natl Acad Sci U S A

journal_title

Proceedings of the National Academy of Sciences of the United States of America

authors

Zhang Z,Tao Z,Gameiro A,Barcelona S,Braams S,Rauen T,Grewer C

doi

10.1073/pnas.0704570104

subject

Has Abstract

pub_date

2007-11-13 00:00:00

pages

18025-30

issue

46

eissn

0027-8424

issn

1091-6490

pii

0704570104

journal_volume

104

pub_type

杂志文章

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