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Structures of the M2 channel-lining segments from nicotinic acetylcholine and NMDA receptors by NMR spectroscopy

Nature Structural Biology volume 6, pages 374–379 (1999)Cite this article

Abstract

The structures of functional peptides corresponding to the predicted channel-lining M2 segments of the nicotinic acetylcholine receptor (AChR) and of a glutamate receptor of the NMDA subtype (NMDAR) were determined using solution NMR experiments on micelle samples, and solid-state NMR experiments on bilayer samples. Both M2 segments form straight transmembrane α-helices with no kinks. The AChR M2 peptide inserts in the lipid bilayer at an angle of 12° relative to the bilayer normal, with a rotation about the helix long axis such that the polar residues face the N-terminal side of the membrane, which is assigned to be intracellular. A model built from these solid-state NMR data, and assuming a symmetric pentameric arrangement of M2 helices, results in a funnel-like architecture for the channel, with the wide opening on the N-terminal intracellular side.

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Figure 1: a–h, Single-channel recordings from recombinant M2 peptides in lipid bilayers.
Figure 2: Solution and solid-state 2D NMR spectra of AchR M2 peptides.
Figure 4: Superposition of the backbone heavy atoms for the 10 lowest energy structures of a, AChR M2 and b, NMDAR M2, determined by solution NMR in DPC micelles.
Figure 3: One-dimensional solid-state 15N chemical-shift NMR spectra of M2 peptides in oriented lipid bilayers.
Figure 5: a–c, Top (a) and side (b,c) stereo views of a model of the AChR M2 funnel-like pentameric bundle.

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Acknowledgements

We thank R. Nelson for assistance with molecular modeling and the preparation of Fig. 5. We thank W. Sun and A. Ferrer-Montiel for their participation in the initial experiments. This research was supported by grants from the National Institute of General Medical Sciences to S.J.O and to M.M. This research utilized the Resource for Solid-State NMR of Proteins at the University of Pennsylvania, supported by a grant from the Biomedical Research Technology Program, National Center for Research Resources, National Institutes of Health. F.M.M. was supported by postdoctoral fellowships from the Natural Sciences and Engineering Research Council of Canada and the Medical Research Council of Canada. A.P.V. was supported by a postdoctoral fellowship from FAPESP-Brazil (Fundacao de Amparo a Pesquisa do Estado de Sao Paulo).

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Author notes

  1. F.M. Marassi

    Present address: The Wistar Institute, 3601 Spruce Street, Philadelphia, Pennsylvania, 19104, USA

  2. J.J. Gesell

    Present address: Schering-Plough Research Institute, 2015 Galloping Hill Road, Kenilworth, New Jersey, 07033, USA

  3. A.P. Valente

    Present address: Departmento de Bioquimica Medica–CCS–ICB, Universidade Federal do Rio de Janeiro, Rio de Janeiro–RJ, CEP 21941–590, Brazil

  4. Y. Kim

    Present address: LG Chemical Ltd./Research Park, Taejon, 305-380, Korea

Authors and Affiliations

  1. Department of Chemistry, University of Pennsylvania, Philadelphia, 19014, Pennsylvania, USA

    S.J. Opella, F.M. Marassi, J.J. Gesell, A.P. Valente & Y. Kim

  2. Department of Biology, University of California, San Diego, La Jolla, 92093, California, USA

    M. Oblatt-Montal & M. Montal

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Opella, S., Marassi, F., Gesell, J. et al. Structures of the M2 channel-lining segments from nicotinic acetylcholine and NMDA receptors by NMR spectroscopy. Nat Struct Mol Biol 6, 374–379 (1999). https://doi.org/10.1038/7610

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