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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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.
The authors declare no competing financial interests.
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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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