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Asparagine-mediated self-association of a model transmembrane helix

Nature Structural Biology volume 7, pages 161–166 (2000)Cite this article

Abstract

In membrane proteins, the extent to which polarity, hydrogen bonding, and van der Waals packing interactions of the buried, internal residues direct protein folding and association of transmembrane segments is poorly understood. The energetics associated with these various interactions should differ substantially between membrane versus water-soluble proteins. To help evaluate these energetics, we have altered a water-soluble, two-stranded coiled-coil peptide to render its sequence soluble in membranes. The membrane-soluble peptide associates in a monomer-dimer-trimer equilibrium, in which the trimer predominates at the highest peptide/detergent ratios. The oligomers are stabilized by a buried Asn side chain. Mutation of this Asn to Val essentially eliminates oligomerization of the membrane-soluble peptide. Thus, within a membrane-like environment, interactions involving a polar Asn side chain provide a strong thermodynamic driving force for membrane helix association.

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Figure 1: Sequences of peptides and SDS-PAGE of MS1 and variants.
Figure 2: Association of MS1 and N14V in C12E8 micelles, as determined by fluorescence resonance energy transfer.
Figure 3: Determination of the association state of MS1 using fluorescence resonance energy transfer in C14-betaine micelles.
Figure 4: Analytical ultracentrifugation of NBD-MS1 in C14-betaine micelles.
Figure 5: Analytical ultracentrifugation of NBD–N14V in C14-betaine micelles.

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Author notes

  1. Christin Choma

    Present address: Department of Chemistry, Rensselaer Polytechnic Institute, Troy, New York, 12180-3590, USA

Authors and Affiliations

  1. Department of Biochemistry and Biophysics, University of Pennsylvania, School of Medicine, Pennsylvania, Philadelphia, 19104-6059, USA

    Christin Choma, Holly Gratkowski, James D. Lear & William F. DeGrado

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  1. Christin Choma
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  4. William F. DeGrado
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Correspondence to James D. Lear or William F. DeGrado.

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Choma, C., Gratkowski, H., Lear, J. et al. Asparagine-mediated self-association of a model transmembrane helix. Nat Struct Mol Biol 7, 161–166 (2000). https://doi.org/10.1038/72440

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