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Interhelical hydrogen bonding drives strong interactions in membrane proteins

Nature Structural Biology volume 7, pages 154–160 (2000)Cite this article

Abstract

Polar residues in transmembrane α-helices may strongly influence the folding or association of integral membrane proteins. To test whether a motif that promotes helix association in a soluble protein could do the same within a membrane, we designed a model transmembrane helix based on the GCN4 leucine zipper. We found in both detergent miscelles and biological membranes that helix association is driven strongly by asparagine, independent of the rest of the hydrophobic leucine and/or valine sequence. Hydrogen bonding between membrane helices gives stronger associations than the packing of surfaces in glycophorin A helices, creating an opportunity to stabilize structures, but also implying a danger that non-specific interactions might occur. Thus, membrane proteins may fold to avoid exposure of strongly hydrogen bonding groups at their lipid exposed surfaces.

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Figure 1: Transmembrane helix designs.
Figure 2: SDS-PAGE of SN chimeras.
Figure 3: Competition experiment with VNVV peptide.
Figure 4: Interactions between SN chimeras and VNVV peptide.
Figure 5: TOXCAT assays of transmembrane helix association.
Figure 6: Proton NMR spectra of the synthetic membrane leucine zipper.
Figure 7: CD of the synthetic membrane peptide VNVV in SDS micelles.
Figure 8: Possible models that describe the equilibrium and hydrogen bonding patterns predicted by the NMR data.

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Acknowledgements

We thank P.D. Adams, K. R. MacKenzie, A. Senes and I. Ubarretxena for helpful discussions. We are also indebted to L. Fisher for advice and assistance in peptide synthesis. We kindly acknowledge G. King for permission to adapt a figure. This research is funded with a program project grant on helix interactions in membrane proteins (National Institute of Health) and the National Foundation for Cancer Research.

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Authors and Affiliations

  1. Department of Molecular Biophysics and Biochemistry, Yale University, 266 Whitney Avenue, P.O. Box 208114, New Haven, 06520-8114, Conneticut, USA

    Fang Xiao Zhou, Melanie J. Cocco, William P. Russ, Axel T. Brunger & Donald M. Engelman

  2. Howard Hughes Medical Institute.,

    Axel T. Brunger

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  1. Fang Xiao Zhou
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Correspondence to Donald M. Engelman.

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Xiao Zhou, F., Cocco, M., Russ, W. et al. Interhelical hydrogen bonding drives strong interactions in membrane proteins. Nat Struct Mol Biol 7, 154–160 (2000). https://doi.org/10.1038/72430

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