Abstract
Neurotransmitter-gated channels, such as the NMDA receptor, are oligomeric membrane proteins assembled from multiple homologous subunits organized around a central ion-conducting pore (1,2). A major effort is directed toward understanding the structural features that determine their function. However, the absence of an atomic resolution structure for even a single ligand-gated ion channel remains a serious deficiency to accomplish this goal. There has been little success in the preparation of high-quality three-dimensional crystals of these membrane proteins, and the determination of their structures in solution by multidimensional NMR spectroscopy has been hampered by the large size and multiplicity of their subunits as well as the requirement of a lipid environment to maintain the native folded conformation. These significant limitations in the conventional methods of structural biology have prompted the development of indirect strategies to identify and characterize those segments of these proteins that contribute directly to the formation of the channel lining. In this context, a biomimicry-based strategy founded on the tenet that a specific functional feature may be transferred to a receptor lacking it by implanting the structural determinants that define the property is contributing considerable diagnostic information. The uniqueness of the biomimicry-based strategy resides in the notion of achieving a “gain of function” in contrast with the classical mutagenesis approach that randomly aims for a “loss of function.”
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References
Sutcliffe, M., Wo, G. Z., and Oswald, R. E. (1996) Three-dimensional models of non-NMDA glutamate receptors. Biophys. J. 70, 1575–1589.
Montal, M. (1995) Design of molecular function: channels of communication. Annu. Rev. Biophys. Biomol. Struct. 24, 31–57.
Galzi, J. L., Devillers-Thiery, A., Hussy, N., Bertrand, S., Changeaux, J. P., and Bertrand, D. (1992) Mutations in the channel domain of a neuronal nicotinic receptor convert ion selectivity from cationic to anionic. Nature 359, 500–505.
Gross, A., Abramson, T., and MacKinnon, R. (1994) Transfer of the scorpion toxin receptor to an insensitive potassium channel. Neuron 13, 961–966.
Ferrer-Montiel, A. V., Sun, W., and Montal, M. (1995) Molecular design of the N-methyl-d-aspartate receptor binding site for phencyclidine and dizolcipine. Proc. Natl. Acad. Sci. USA 92, 8021–8025.
Zarei, M. M. and Dani, J. A. (1995) Structural basis for explaining open-channel blockade of the NMDA receptor. J. Neurosci. 15, 1446–1454.
McBurney, R. W. (1997) Development of the NMDA ion-channel blocker, aptiganel hydrochloride, as a neuroprotective agent for acute CNS injury. Int. Rev. Neurobiol. 40, 173–195.
Ferrer-Montiel, A. V., Sun, W., and Montal, M. (1996) A single tryptophan on M2 of glutamate receptor channels confers high permeability to divalent cations. Biophys. J. 71, 749–758.
Kuner, T., Wollmuth, L. P., Karlin, A., Seeburg, P. H., and Sackman, B. (1996) Structure of the NMDA receptor channel M2 segment inferred from the accessibility of substituted cysteines. Neuron 17, 343–352.
Nakanishi, S. and Masu, M. (1994) Molecular diversity and functions of glutamate receptors. Annu. Rev. Biophys. Biomol. Struct. 23, 319–348.
Hollman, M. and Heinemann, S. F. (1994). Cloned glutamate receptors. Annu. Rev. Neurosci. 17, 31–108.
Burnashev, N., Schoepfer, R., Monyer, H., Ruppersberg, J. P., Gunther, W., Seeburg, P. H., et al. (1992) Control by asparagine residues of calcium permeability and magnesium blockade in the NMDA receptor. Science 257, 1415–1419.
Burnashev, N., Monyer, H., Seeburg, P. H., and Sakmann, B. (1992) Divalent ion permeability of AMPA receptor channels is dominated by the edited form of a single subunit. Neuron 8, 189–198.
Dingledine, R., Hume, R. I., and Heinemann, S. F. (1992) Structural determinants of barium permeation and rectification in non-NMDA receptor channels. J. Neurosci. 12, 4080–4087.
Sakurada, K., Masu, M., and Nakanishi (1993) Alteration of Ca2+ permeability and sensitivity to Mg2+ and channel blockers by a single amino acid substitution in the N-methyl-d-aspartate receptor. J. Biol. Chem. 268, 410–415.
Verdoorn, T. A., Burnashev, N., Monyer, H., Seeburg, P. H., and Sakman, B. (1991) Structural determinants of ion flow through recombinant glutamate receptor channels. Science 252, 1715–1718.
Egebjerg, J. and Heinemann, S. F. (1993) Ca2+ permeability of unedited and edited versions of the kainate selective glutamate receptor GluR6. Proc. Natl. Acad. Sci. USA 90, 755–759.
Ferrer-Montiel, A. V. and Montal, M. (1994) Structure-function relations in ligand-gated ion channels: reconstitution in lipid bilayers and heterologous expression in Xenopus oocytes. Methods: Companion Methods Enzymol. 6, 60–69.
Ferrer-Montiel, A. V., Merino, J. M., Planells-Cases, R., Sun, W., and Montel, M. (1998) Structural determinants of the blocker binding site in glutamate and NMDA receptor channels. Neuropharmacology 37, 139–147.
Ferrer-Montiel, A. V. and Montal, M. (1996) Pentameric subunit stoichiometry of a neuronal glutamate receptor. Proc. Natl. Acad. Sci. USA 93, 2741–2744.
Premkumar, L. S. and Auerbach, A. (1996) Stoichiometry of recombinant NMDA receptor channels. Soc. Neurosci. Abstracts 22, 593; Abstract no. 237.8.
Mayer, M. L. and Westbrook, G. L. (1987) Permeation and block of N-methyl-d-aspartic acid receptor channels by divalent cations in mouse cultured central neurons. J. Physiol. (Lond.) 286, 417–445.
Hille, B. (1992) Ion Channels of Excitable Cells, 2nd ed., Sinauer Assoc., Sunderland, MA.
Miledi, R. and Parker, I. (1984) Chloride current induced by injection of calcium into Xenopus oocytes. J. Physiol. (Lond.) 357, 173–188.
Woodhull, A. M. (1973) Ionic blockage of sodium channels in nerve. J. Gen Physiol. 61, 687–708.
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Ferrer-Montiel, A.V., Montal, M. (1999). Engineering the NMDA Receptor Channel Lining. In: Li, M. (eds) NMDA Receptor Protocols. Methods in Molecular Biology™, vol 128. Humana Press. https://doi.org/10.1385/1-59259-683-5:167
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DOI: https://doi.org/10.1385/1-59259-683-5:167
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