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Science 1 August 2003:
Vol. 301. no. 5633, pp. 616 - 620
DOI: 10.1126/science.1087619
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Research Articles

Structure and Mechanism of the Glycerol-3-Phosphate Transporter from Escherichia coli

Yafei Huang,* M. Joanne Lemieux,*{dagger} Jinmei Song, Manfred Auer, Da-Neng Wang{ddagger}

The major facilitator superfamily represents the largest group of secondary membrane transporters in the cell. Here we report the 3.3 angstrom resolution structure of a member of this superfamily, GlpT, which transports glycerol-3-phosphate into the cytoplasm and inorganic phosphate into the periplasm. The amino- and carboxyl-terminal halves of the protein exhibit a pseudo two-fold symmetry. Closed off to the periplasm, a centrally located substrate-translocation pore contains two arginines at its closed end, which comprise the substrate-binding site. Upon substrate binding, the protein adopts a more compact conformation. We propose that GlpT operates by a single–binding site, alternating-access mechanism through a rocker-switch type of movement.

Skirball Institute of Biomolecular Medicine and Department of Cell Biology, New York University School of Medicine, 540 First Avenue, New York, NY 10016, USA.



* These authors contributed equally to this work.

{dagger} Present address: Department of Biochemistry, University of Alberta, Edmonton, Canada T6G 2H6.

{ddagger} To whom correspondence should be addressed. E-mail: wang{at}saturn.med.nyu.edu

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A Member of the Sugar Transporter Family, Stl1p Is the Glycerol/H+ Symporter in Saccharomyces cerevisiae.
C. Ferreira, F. van Voorst, A. Martins, L. Neves, R. Oliveira, M. C. Kielland-Brandt, C. Lucas, and A. Brandt (2005)
Mol. Biol. Cell 16, 2068-2076
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Identification of Essential Amino Acid Residues of the NorM Na+/Multidrug Antiporter in Vibrio parahaemolyticus.
M. Otsuka, M. Yasuda, Y. Morita, C. Otsuka, T. Tsuchiya, H. Omote, and Y. Moriyama (2005)
J. Bacteriol. 187, 1552-1558
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Modeling membrane proteins based on low-resolution electron microscopy maps: a template for the TM domains of the oxalate transporter OxlT.
T. Beuming and H. Weinstein (2005)
Protein Eng. Des. Sel. 18, 119-125
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Missense Mutations That Inactivate the Aspergillus nidulans nrtA Gene Encoding a High-Affinity Nitrate Transporter.
J. R. Kinghorn, J. Sloan, G. J. M. Kana'n, E. R. DaSilva, D. A. Rouch, and S. E. Unkles (2005)
Genetics 169, 1369-1377
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Small-Scale Molecular Motions Accomplish Glutamate Uptake in Human Glutamate Transporters.
H. P. Koch and H. P. Larsson (2005)
J. Neurosci. 25, 1730-1736
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Altered Oxyanion Selectivity in Mutants of UhpT, the Pi -linked Sugar Phosphate Carrier of Escherichia coli.
J. A. Hall and P. C. Maloney (2005)
J. Biol. Chem. 280, 3376-3381
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Promiscuity in the Geometry of Electrostatic Interactions between the Escherichia coli Multidrug Resistance Transporter MdfA and Cationic Substrates.
J. Adler and E. Bibi (2005)
J. Biol. Chem. 280, 2721-2729
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