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Crystal structure of a catalytic intermediate of the maltose transporter

Abstract

The maltose uptake system of Escherichia coli is a well-characterized member of the ATP-binding cassette transporter superfamily. Here we present the 2.8-Å crystal structure of the intact maltose transporter in complex with the maltose-binding protein, maltose and ATP. This structure, stabilized by a mutation that prevents ATP hydrolysis, captures the ATP-binding cassette dimer in a closed, ATP-bound conformation. Maltose is occluded within a solvent-filled cavity at the interface of the two transmembrane subunits, about halfway into the lipid bilayer. The binding protein docks onto the entrance of the cavity in an open conformation and serves as a cap to ensure unidirectional translocation of the sugar molecule. These results provide direct evidence for a concerted mechanism of transport in which solute is transferred from the binding protein to the transmembrane subunits when the cassette dimer closes to hydrolyse ATP.

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Figure 1: Structure of the maltose transporter in a catalytic intermediate conformation.
Figure 2: Ribbon diagram of the MalK subunits with bound ATP.
Figure 3: Architecture of the transmembrane subunits.
Figure 4: The TMD–MalK interface.
Figure 5: Transfer of the maltose from MBP to the TM binding site.
Figure 6: Comparison of the structures of two binding-protein-dependent ABC uptake systems.
Figure 7: A model for the ABC uptake system.

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Acknowledgements

We thank the beamline staff at the Advanced Photon Source beamline 23-ID for assistance with data collection, and R. MacKinnon, H. Shuman, D. Yernool and C. Orelle for discussions. This work was supported by NIH grants (J.C., A.L.D. and F.A.Q.), the Welch Foundation (F.A.Q.), the Pew Scholar Program (J.C.) and a postdoctoral fellowship from American Heart Association (M.L.O).

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Oldham, M., Khare, D., Quiocho, F. et al. Crystal structure of a catalytic intermediate of the maltose transporter. Nature 450, 515–521 (2007). https://doi.org/10.1038/nature06264

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