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Carbon catabolite repression of the maltose transporter revealed by X-ray crystallography

Abstract

Efficient carbon utilization is critical to the survival of microorganisms in competitive environments. To optimize energy usage, bacteria have developed an integrated control system to preferentially uptake carbohydrates that support rapid growth. The availability of a preferred carbon source, such as glucose, represses the synthesis and activities of proteins necessary for the transport and metabolism of secondary carbon sources. This regulatory phenomenon is defined as carbon catabolite repression1. In enteric bacteria, the key player of carbon catabolite repression is a component of the glucose-specific phosphotransferase system, enzyme IIA (EIIAGlc)1,2. It is known that unphosphorylated EIIAGlc binds to and inhibits a variety of transporters when glucose is available1,2. However, understanding the underlying molecular mechanism has been hindered by the complete absence of structures for any EIIAGlc–transporter complexes. Here we present the 3.9 Å crystal structure of Escherichia coli EIIAGlc in complex with the maltose transporter, an ATP-binding cassette (ABC) transporter. The structure shows that two EIIAGlc molecules bind to the cytoplasmic ATPase subunits, stabilizing the transporter in an inward-facing conformation and preventing the structural rearrangements necessary for ATP hydrolysis. We also show that the half-maximal inhibitory concentrations of the full-length EIIAGlc and an amino-terminal truncation mutant differ by 60-fold, consistent with the hypothesis that the amino-terminal region, disordered in the crystal structure, functions as a membrane anchor to increase the effective EIIAGlc concentration at the membrane3,4. Together these data suggest a model of how the central regulatory protein EIIAGlc allosterically inhibits maltose uptake in E. coli.

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Figure 1: Two orthogonal views of the EIIAGlc–MalFGK2 complex.
Figure 2: Binding of EIIAGlc prevents MalK closure.
Figure 3: The N terminus of EIIAGlc probably functions as a membrane anchor.
Figure 4: Inhibition of the maltose transporter by inducer exclusion.

Accession codes

Accessions

Protein Data Bank

Data deposits

Coordinates and structure factors have been deposited in the Protein Data Bank under accession number 4JBW.

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Acknowledgements

We thank the staff at the Advance Photon Source GM/CA-CAT, NE-CAT and SBC for assistance with data collection. This work was supported by a National Institutes of Health grant (GM070515 to J.C. and A.L.D.).

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All authors designed the study and analysed the data. S.C. crystallized the complex and performed the biochemical experiments. S.C. and M.L.O. determined the crystal structure and made the figures. J.C. wrote the manuscript with inputs from all authors.

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Correspondence to Jue Chen.

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Chen, S., Oldham, M., Davidson, A. et al. Carbon catabolite repression of the maltose transporter revealed by X-ray crystallography. Nature 499, 364–368 (2013). https://doi.org/10.1038/nature12232

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