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The dynamics of the G protein-coupled neuropeptide Y2 receptor in monounsaturated membranes investigated by solid-state NMR spectroscopy

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Abstract

In contrast to the static snapshots provided by protein crystallography, G protein-coupled receptors constitute a group of proteins with highly dynamic properties, which are required in the receptors’ function as signaling molecule. Here, the human neuropeptide Y2 receptor was reconstituted into a model membrane composed of monounsaturated phospholipids and solid-state NMR was used to characterize its dynamics. Qualitative static 15N NMR spectra and quantitative determination of 1H–13C order parameters through measurement of the 1H–13C dipolar couplings of the CH, CH2 and CH3 groups revealed axially symmetric motions of the whole molecule in the membrane and molecular fluctuations of varying amplitude from all molecular segments. The molecular order parameters (Sbackbone = 0.59–0.67, SCH2 = 0.41–0.51 and SCH3 = 0.22) obtained in directly polarized 13C NMR experiments demonstrate that the Y2 receptor is highly mobile in the native-like membrane. Interestingly, according to these results the receptor was found to be slightly more rigid in the membranes formed by the monounsaturated phospholipids than by saturated phospholipids as investigated previously. This could be caused by an increased chain length of the monounsaturated lipids, which may result in a higher helical content of the receptor. Furthermore, the incorporation of cholesterol, phosphatidylethanolamine, or negatively charged phosphatidylserine into the membrane did not have a significant influence on the molecular mobility of the Y2 receptor.

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Acknowledgments

D. H. would like to acknowledge valuable discussions with Profs. D. Reichert and P. K. Madhu. Part of this study was supported by the Europäischer Sozialfonds (ESF 22117016 and 24127009). We would like to thank the group of Annette G. Beck-Sickinger for the TAMRA labeled NPY.

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Authors and Affiliations

  1. Institute of Medical Physics and Biophysics, University of Leipzig, Härtelstr. 16-18, 04107, Leipzig, Germany

    Lars Thomas, Julian Kahr, Peter Schmidt, Ulrike Krug, Holger A. Scheidt & Daniel Huster

  2. University of Applied Sciences, Dr.-Friedrichs-Ring 2A, 08056, Zwickau, Germany

    Julian Kahr

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  1. Lars Thomas
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  2. Julian Kahr
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  3. Peter Schmidt
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Correspondence to Daniel Huster.

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Lars Thomas and Julian Kahr have contributed equally to this work.

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Thomas, L., Kahr, J., Schmidt, P. et al. The dynamics of the G protein-coupled neuropeptide Y2 receptor in monounsaturated membranes investigated by solid-state NMR spectroscopy. J Biomol NMR 61, 347–359 (2015). https://doi.org/10.1007/s10858-014-9892-5

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