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Crystal structure of a phosphorylation-coupled saccharide transporter

Nature volume 473, pages 50–54 (2011)Cite this article

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Abstract

Saccharides have a central role in the nutrition of all living organisms. Whereas several saccharide uptake systems are shared between the different phylogenetic kingdoms, the phosphoenolpyruvate-dependent phosphotransferase system exists almost exclusively in bacteria. This multi-component system includes an integral membrane protein EIIC that transports saccharides and assists in their phosphorylation. Here we present the crystal structure of an EIIC from Bacillus cereus that transports diacetylchitobiose. The EIIC is a homodimer, with an expansive interface formed between the amino-terminal halves of the two protomers. The carboxy-terminal half of each protomer has a large binding pocket that contains a diacetylchitobiose, which is occluded from both sides of the membrane with its site of phosphorylation near the conserved His 250 and Glu 334 residues. The structure shows the architecture of this important class of transporters, identifies the determinants of substrate binding and phosphorylation, and provides a framework for understanding the mechanism of sugar translocation.

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Figure 1: Function and structure of ChbC.
Figure 2: The C-terminal sugar-binding domain.
Figure 3: Proposed conformational changes in sugar transport.

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Protein Data Bank

Data deposits

Atomic coordinates and structure factors have been deposited with the Protein Data Bank under accession code 3QNQ.

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Acknowledgements

Data for this study were measured at beamlines X4A, X4C, X25 and X29 of the National Synchrotron Light Source and the NE-CAT 24ID-C and E at the Advanced Photon Source. This work was supported by the US National Institutes of Health (DK088057, GM098878 and GM05026-sub0007 to M.Z., and T32HL087745 to E.J.L.). M.Z. is a Pew Scholar in Biomedical Sciences. The NYCOMPS central facility was supported by GM05026 to W.A.H. as part of the Protein Structure Initiative (PSI-2) established by the National Institute of General Medical Sciences. The authors would like to thank B. Honig for support and M. Saier, B. Erni, R. Kaback and D.-N. Wang for comments on the manuscript and helpful discussions. M.Z. is grateful to R. MacKinnon for advice and encouragement.

Author information

Author notes

  1. Xiangshu Jin, Elena J. Levin, Hua Huang and Yinong Zong: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Physiology & Cellular Biophysics, College of Physicians and Surgeons, Columbia University, 630 West 168th Street, New York, 10032, New York, USA

    Yu Cao, Elena J. Levin, Hua Huang, Jun Weng, Yaping Pan & Ming Zhou

  2. Department of Biochemistry and Molecular Biophysics, Center for Computational Biology and Bioinformatics, Howard Hughes Medical Institute, Columbia University, 1130 St. Nicholas Ave, Room 815, New York, 10032, New York, USA

    Xiangshu Jin

  3. Sanford-Burnham Institute, La Jolla, 92037, California, USA

    Yinong Zong

  4. Department of Psychiatry and Center for Molecular Recognition, Columbia University, 630 West 168th Street, New York, 10032, New York, USA

    Matthias Quick & Jonathan A. Javitch

  5. Division of Molecular Therapeutics, New York State Psychiatric Institute, 1051 Riverside Drive, New York, 10032, New York, USA

    Matthias Quick & Jonathan A. Javitch

  6. New York Consortium on Membrane Protein Structure, New York Structural Biology Center, 89 Convent Avenue, New York, 10027, New York, USA

    James Love, Marco Punta, Burkhard Rost & Wayne A. Hendrickson

  7. Department of Computer Science and Institute for Advanced Study, Technical University of Munich, D-85748 Munich, Germany

    Marco Punta & Burkhard Rost

  8. Department of Biochemistry and Molecular Biophysics, Howard Hughes Medical Institute, Columbia University, 630 West 168th Street, New York, 10032, New York, USA

    Wayne A. Hendrickson

  9. Department of Pharmacology, Columbia University, 630 West 168th Street, New York, 10032, New York, USA

    Jonathan A. Javitch

  10. Department of Chemistry and Chemical Biology, Cornell University, NE-CAT, Advanced Photon Source, Argonne, 60439, Illinois, USA

    Kanagalaghatta R. Rajashankar

Authors
  1. Yu Cao
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Contributions

J.L., M.P., B.R. and W.A.H. identified ChbC homologues in the database. J.L. carried out the cloning and the initial expression studies. Y.C., H.H., E.J.L., J.W. and M.Z. performed protein expression, purification, crystallization and X-ray diffraction data collection and analysis. X.J., Y.Z., E.J.L. and M.Z. solved and refined the structures. M.Q., Y.C., Y.P., J.A.J. and M.Z. characterized ChbC function. K.R.R. advised on data collection and crystallography. E.J.L. and M.Z. wrote the manuscript with input from all authors.

Corresponding author

Correspondence to Ming Zhou.

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Cao, Y., Jin, X., Levin, E. et al. Crystal structure of a phosphorylation-coupled saccharide transporter. Nature 473, 50–54 (2011). https://doi.org/10.1038/nature09939

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