Pentameric assembly of a neuronal glutamate transporter

Sepehr Eskandari, Michael Kreman, Michael P. Kavanaugh, Ernest M. Wright, Guido A. Zampighi

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109 Scopus citations


Freeze-fracture electron microscopy was used to study the structure of a human neuronal glutamate transporter (EAAT3). EAAT3 was expressed in Xenopus laevis oocytes, and its function was correlated with the total number of transporters in the plasma membrane of the same cells. Function was assayed as the maximum charge moved in response to a series of transmembrane voltage pulses. The number of transporters in the plasma membrane was determined from the density of a distinct 10-nm freeze-fracture particle, which appeared in the protoplasmic face only after EAAT3 expression. The linear correlation between EAAT3 maximum carrier-mediated charge and the total number of the 10-nm particles suggested that this particle represented functional EAAT3 in the plasma membrane. The cross-sectional area of EAAT3 in the plasma membrane (48 ± 5 nm2) predicted 35 ± 3 transmembrane α-helices in the transporter complex. This information along with secondary structure models (6-10 transmembrane α-helices) suggested an oligomeric state for EAAT3. EAAT3 particles were pentagonal in shape in which five domains could be identified. They exhibited five-fold symmetry because they appeared as equilateral pentagons and the angle at the vertices was 110°. Each domain appeared to contribute to an extracellular mass that projects ≃3 nm into the extracellular space. Projections from all five domains taper toward an axis passing through the center of the pentagon, giving the transporter complex the appearance of a penton-based pyramid. The pentameric structure of EAAT3 offers new insights into its function as both a glutamate transporter and a glutamate-gated chloride channel.

Original languageEnglish
Pages (from-to)8641-8646
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Issue number15
StatePublished - Jul 18 2000


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