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X-ray structure of the Yersinia pestis heme transporter HmuUV

Nature Structural & Molecular Biology volume 19, pages 1310–1315 (2012)Cite this article

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Abstract

HmuUV is a bacterial ATP-binding cassette (ABC) transporter that catalyzes heme uptake into the cytoplasm of the Gram-negative pathogen Yersinia pestis. We report the crystal structure of HmuUV at 3.0 Å resolution in a nucleotide-free state, which features a heme translocation pathway in an outward-facing conformation, poised to accept a heme from the cognate periplasmic binding protein HmuT. A new assay allowed us to determine in vitro rates of HmuUV-catalyzed heme transport into proteoliposomes and to establish the role of conserved residues in the translocation pathway of HmuUV and at the interface with HmuT. Differences in architecture relative to the related vitamin B12 transporter BtuCD suggest an adaptation of HmuUV for its smaller substrate. Our study also suggests that type II ABC importers, which include bacterial iron-siderophore, heme and cobalamin transporters, have a coupling mechanism distinct from that of other ABC transporters.

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Figure 1: In vitro heme transport assay.
Figure 2: Structure of Y. pestis HmuUV in ribbon representation.
Figure 3: Conformation of the TMDs and sequence conservation in the gating helices.
Figure 4: Sequence conservation in the gating helices.
Figure 5: Function of conserved residues in the gating helices.

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Acknowledgements

We thank the beamline staff at the Swiss Light Source for assistance with data collection, R.D. Perry (University of Kentucky, Lexington) for providing the DNA encoding the Y. pestis HmuUV–T system. This research was supported by the National Center for Competence in Research Structural Biology Zurich and the Swiss National Science Foundation (grant SNF 31003A-131075/1 to K.P.L.).

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  1. Jae-Sung Woo and Antra Zeltina: These authors contributed equally to this work.

Authors and Affiliations

  1. Institute of Molecular Biology and Biophysics, Eidgenössische Technische Hochschule (ETH) Zürich, Zürich, CH-8093, Switzerland

    Jae-Sung Woo, Antra Zeltina, Birke A Goetz & Kaspar P Locher

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  1. Jae-Sung Woo
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  2. Antra Zeltina
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Contributions

B.A.G. performed protein purification and crystallization, K.P.L. determined the crystal structure and built the initial model, J.-S.W. performed structure refinement and bioinformatics analysis, A.Z. performed functional analysis, J.-S.W., A.Z. and K.P.L. analyzed the data and wrote the manuscript.

Corresponding author

Correspondence to Kaspar P Locher.

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The authors declare no competing financial interests.

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Woo, JS., Zeltina, A., Goetz, B. et al. X-ray structure of the Yersinia pestis heme transporter HmuUV. Nat Struct Mol Biol 19, 1310–1315 (2012). https://doi.org/10.1038/nsmb.2417

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