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Transmembrane helices containing a charged arginine are thermodynamically stable

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Abstract

Hydrophobic amino acids are abundant in transmembrane (TM) helices of membrane proteins. Charged residues are sparse, apparently due to the unfavorable energetic cost of partitioning charges into nonpolar phases. Nevertheless, conserved arginine residues within TM helices regulate vital functions, such as ion channel voltage gating and integrin receptor inactivation. The energetic cost of arginine in various positions along hydrophobic helices has been controversial. Potential of mean force (PMF) calculations from atomistic molecular dynamics simulations predict very large energetic penalties, while in vitro experiments with Sec61 translocons indicate much smaller penalties, even for arginine in the center of hydrophobic TM helices. Resolution of this conflict has proved difficult, because the in vitro assay utilizes the complex Sec61 translocon, while the PMF calculations rely on the choice of simulation system and reaction coordinate. Here we present the results of computational and experimental studies that permit direct comparison with the Sec61 translocon results. We find that the Sec61 translocon mediates less efficient membrane insertion of Arg-containing TM helices compared with our computational and experimental bilayer-insertion results. In the simulations, a combination of arginine snorkeling, bilayer deformation, and peptide tilting is sufficient to lower the penalty of Arg insertion to an extent such that a hydrophobic TM helix with a central Arg residue readily inserts into a model membrane. Less favorable insertion by the translocon may be due to the decreased fluidity of the endoplasmic reticulum (ER) membrane compared with pure palmitoyloleoyl-phosphocholine (POPC). Nevertheless, our results provide an explanation for the differences between PMF- and experiment-based penalties for Arg burial.

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Acknowledgements

This research was supported by a Marie Curie International Fellowship to M.B.U., grants from the National Institute of General Medical Science GM74737 (S.H.W.), Program Project GM86685 from NINDS and NIGMS (S.H.W., D.J.T.), NSF grant CHE-0750175 (D.J.T.), and from the European Research Council (ERC-2008-AdG 232648), the Swedish Cancer Foundation, the Swedish Research Council, and the Swedish Foundation for Strategic Research (G.v.H.).

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Authors and Affiliations

  1. Institute for NanoBioTechnology and Department of Materials Science, Johns Hopkins University, Baltimore, MD, 21218, USA

    Martin B. Ulmschneider

  2. Institute of Natural Sciences, Shanghai Jiao Tong University, Shanghai, 200240, China

    Jakob P. Ulmschneider

  3. Department of Chemistry and the Center for Biomembrane Systems, University of California, Irvine, CA, 92697-2025, USA

    J. Alfredo Freites & Douglas J. Tobias

  4. Department of Biochemistry and Biophysics, Stockholm University, SE-106 91, Stockholm, Sweden

    Gunnar von Heijne

  5. Department of Physiology and Biophysics and the Center for Biomembrane Systems, University of California, Irvine, CA, 92697-4560, USA

    Stephen H. White

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  1. Martin B. Ulmschneider
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  2. Jakob P. Ulmschneider
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  3. J. Alfredo Freites
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Correspondence to Stephen H. White.

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“Special Issue: Shining Light on Membrane Proteins”

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Ulmschneider, M.B., Ulmschneider, J.P., Freites, J.A. et al. Transmembrane helices containing a charged arginine are thermodynamically stable. Eur Biophys J 46, 627–637 (2017). https://doi.org/10.1007/s00249-017-1206-x

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  • DOI: https://doi.org/10.1007/s00249-017-1206-x

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