Abstract
We report a structure of a trimeric glutamate transporter homolog from Pyrococcus horikoshii with two protomers in an inward facing state and the third in an intermediate conformation between the outward and inward facing states. The intermediate shows a cavity in the thinnest region of the transporter, which is potentially accessible to extracellular and cytoplasmic solutions. Our findings suggest a structural principle by which transport intermediates may mediate uncoupled permeation of polar solutes.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$189.00 per year
only $15.75 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Danbolt, N.C. Prog. Neurobiol. 65, 1–105 (2001).
Fairman, W.A., Vandenberg, R.J., Arriza, J.L., Kavanaugh, M.P. & Amara, S.G. Nature 375, 599–603 (1995).
Wadiche, J.I., Arriza, J.L., Amara, S.G. & Kavanaugh, M.P. Neuron 14, 1019–1027 (1995).
Arriza, J.L., Eliasof, S., Kavanaugh, M.P. & Amara, S.G. Proc. Natl. Acad. Sci. USA 94, 4155–4160 (1997).
Veruki, M.L., Morkve, S.H. & Hartveit, E. Nat. Neurosci. 9, 1388–1396 (2006).
Yernool, D., Boudker, O., Jin, Y. & Gouaux, E. Nature 431, 811–818 (2004).
Boudker, O., Ryan, R.M., Yernool, D., Shimamoto, K. & Gouaux, E. Nature 445, 387–393 (2007).
Reyes, N., Ginter, C. & Boudker, O. Nature 462, 880–885 (2009).
Poulsen, M.V. & Vandenberg, R.J. J. Physiol. (Lond.) 534, 159–167 (2001).
Zerangue, N., Arriza, J.L., Amara, S.G. & Kavanaugh, M.P. J. Biol. Chem. 270, 6433–6435 (1995).
Fairman, W.A., Sonders, M.S., Murdoch, G.H. & Amara, S.G. Nat. Neurosci. 1, 105–113 (1998).
Ryan, R.M., Mitrovic, A.D. & Vandenberg, R.J. J. Biol. Chem. 279, 20742–20751 (2004).
Ryan, R.M. & Mindell, J.A. Nat. Struct. Mol. Biol. 14, 365–371 (2007).
Huang, S. & Vandenberg, R.J. Biochemistry 46, 9685–9692 (2007).
Seal, R.P., Shigeri, Y., Eliasof, S., Leighton, B.H. & Amara, S.G. Proc. Natl. Acad. Sci. USA 98, 15324–15329 (2001).
Borre, L., Kavanaugh, M.P. & Kanner, B.I. J. Biol. Chem. 277, 13501–13507 (2002).
Ryan, R.M. & Vandenberg, R.J. J. Biol. Chem. 277, 13494–13500 (2002).
Shachnai, L., Shimamoto, K. & Kanner, B.I. Neuropharmacology 49, 862–871 (2005).
Borre, L. & Kanner, B.I. J. Biol. Chem. 276, 40396–40401 (2001).
Vandenberg, R.J., Huang, S. & Ryan, R.M. Channels (Austin) 2, 51–58 (2008).
Acknowledgements
We thank N. Reyes (Pasteur Institute) for assistance with the design of isothermal titration calorimetry experiments and helpful discussions. V. Chaptal (Institute of Biology and Chemistry of Proteins) is acknowledged for his help with the anisotropy correction procedure and M. Sawaya (Institute for Genomics and Proteomics, University of California, Los Angeles) for providing the suite of programs used for the anisotropy correction. We are thankful for access to beamlines X25 and X29 at the National Synchrotron Light Source. This work was supported by the National Institute of Neurological Disorders and Stroke, grant no. NS064357 to O.B.
Author information
Authors and Affiliations
Contributions
G.V. and O.B. designed the experiments, analyzed data and wrote the manuscript. G.V. conducted the experiments and collected X-ray data.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary Text and Figures
Supplementary Figures 1–11, Supplementary Tables 1 and 2, Supplementary Discussion and Supplementary Methods (PDF 1620 kb)
Rights and permissions
About this article
Cite this article
Verdon, G., Boudker, O. Crystal structure of an asymmetric trimer of a bacterial glutamate transporter homolog. Nat Struct Mol Biol 19, 355–357 (2012). https://doi.org/10.1038/nsmb.2233
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nsmb.2233