Abstract
ATP-binding cassette (ABC) transporters are integral membrane proteins that translocate a diverse array of substrates across cell membranes. We present here the dynamics of complex formation of three structurally characterized ABC transporters—the BtuCD vitamin B12 importer and MetNI d/l-methionine importer from Escherichia coli and the Hi1470/1 metal-chelate importer from Haemophilus influenzae—in complex with their cognate binding proteins. Similarly to other ABC importers, MetNI interacts with its binding protein with low affinity (Kd ∼10−4 M). In contrast, BtuCD–BtuF and Hi1470/1–Hi1472 form stable, high-affinity complexes (Kd ∼10−13 and 10−9 M, respectively). In BtuCD–BtuF, vitamin B12 accelerates the complex dissociation rate ∼107-fold, with ATP having an additional destabilizing effect. The findings presented here highlight substantial mechanistic differences between BtuCD–BtuF, and likely Hi1470/1–Hi1472, and the better-characterized maltose and related ABC transport systems, indicating that there is considerable mechanistic diversity within this large protein super-family.
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Acknowledgements
We thank H. Pinkett for insightful discussions and critical reading of the manuscript and J. Klein and J. Vielmetter for their help in the initial BiaCore experiments. The work was supported in part by US National Institutes of Health grant GM045162, by the Howard Hughes Medical Institute and by fellowships to O.L. from the Fulbright Foundation and the Jane Coffin Childs Memorial Fund for Medical Research.
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A.T.L. and K.P.L. generated the original constructs used in this work; O.L. and D.C.R. designed the research; O.L. and A.T.L. performed the research; O.L., A.T.L., K.P.L. and D.C.R. analyzed the data; and O.L., K.P.L. and D.C.R. wrote the paper.
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Lewinson, O., Lee, A., Locher, K. et al. A distinct mechanism for the ABC transporter BtuCD–BtuF revealed by the dynamics of complex formation. Nat Struct Mol Biol 17, 332–338 (2010). https://doi.org/10.1038/nsmb.1770
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DOI: https://doi.org/10.1038/nsmb.1770
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