Research Article | Published:

Selected peptides targeted to the NMDA receptor channel protect neurons from excitotoxic death

Nature Biotechnology volume 16, pages 286291 (1998) | Download Citation

Subjects

Abstract

Excitotoxic neuronal death, associated with neurodegeneration and stroke, is triggered primarily by massive Ca2+ influx arising from overactivation of glutamate receptor channels of the N-methyl-D-aspartate (NMDA) subtype. To search for channel blockers, synthetic combinatorial libraries were assayed for block of agonist-evoked currents by the human NR1-NR2A NMDA receptor subunits expressed in amphibian oocytes. A set of arginine-rich hexapeptides selectively blocked the NMDA receptor channel with IC50, approximately 100 nM, a potency similar to clinically tolerated blockers such as memantine, and only marginally blocked on non-NMDA glutamate receptors. These peptides prevent neuronal cell death elicited by an excitotoxic insult on hippocampal cultures.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    and 1990. The role of glutamate neurotoxicity in hypoxic-ischemic neuronal death. Annu. Rev. Neurosci. 13: 171–182.

  2. 2.

    1995. Calcium: still center-stage in hypoxic-ischemic neuronal death. Trends Neurosci. 18: 58–60.

  3. 3.

    , , , and 1997. Neuronal necrosis and apoptosis: two distinct events induced by exposure to glutamate or oxidative stress. Adv. Neural. 72: 95–101.

  4. 4.

    1997. Controlling cell death. Science 275: 1081–1082.

  5. 5.

    1996. No NO prevents parkinsonism. Nature Med.. 2: 965–966.

  6. 6.

    and 1996. Nitric oxide synthase: role as a transmitter/mediator in the brain and endocrine system. Annu. Rev. Med. 47: 219–227.

  7. 7.

    , , and 1996. Mitochondrial dysfunction is a primary event in glutamate neurotoxicity. J. Neurosci. 16: 6125–6133.

  8. 8.

    , , , , , et al. 1992. Open channel block of the N-methyl-D-aspartate (NMDA) responses by memantine: Therapeutic advantage against NMDA receptor-mediated neurotoxicity. J. Neurosci. 12: 4427–4436.

  9. 9.

    , , , , , et al. 1991. Generation and use of synthetic peptide combinatorial libraries for basic research and drug delivery. Nature 354: 84–86.

  10. 10.

    , , and 1992. Rapid identification of high affinity peptide ligands using positional scanning synthetic peptide combinatorial libraries. Biotechniques 13: 901–905.

  11. 11.

    , , , , , and 1992. Control by asparagine residues of calcium permeability and magnessium blockade in the NMDA receptor. Science 257: 1415–1419.

  12. 12.

    , , , , , et al. 1986. The anticonvulsant MK-801 is a potent N-methyl-D-aspartate antagonist. Proc. Natl. Acad. Sci. USA 83: 7104–7108.

  13. 13.

    and 1988. Block of N-methyl-D-aspartate-activated current by the anticonvulsant MK-801: selective binding to open channels. Proc Natl. Acad. Sci. USA 85: 1307–1311.

  14. 14.

    and 1996. Identification of a high affinity divalent cation binding site near the entrance of the NMDA receptor channel. Neuron 16: 869–880.

  15. 15.

    , , , , , et al. 1996. Antagonist properties of the stereoisomers of ifenprodil at NR1A/NR2A and NR1A/NR2B subtypes of the NMDA receptor expressed in Xenopus oocytes. Eur. J. Pharmacol. 296: 209–213.

  16. 16.

    , , , , , et al. 1997. In vivo T helper cell response to retro-inverso peptidomimetics. J. Immunol. 159: 3230–3237.

  17. 17.

    et al. 1996. In vitro selection of peptides acting at a new site of NMDA glutamate receptors. Bio/Technology 14: 986–991.

  18. 18.

    , , , , , et al. 1997. Neurotrophic immunophilin ligands stimulate structural and functional recovery in neurodegenerative animal models. Proc. Natl. Acad. Sci. USA 94: 2019–2024.

  19. 19.

    , , , , , et al. 1997. Inhibition of interleukin 1β converting enzyme family proteases reduces ischemic and excitotoxic neuronal damage. Proc. Natl. Acad. Sci. USA 94: 2007–2012.

  20. 20.

    1997. Calcium antagonists: their role in neuroprotection. Int. Rev. Neurobiol. 40: 95–108.

  21. 21.

    1997. GABA and neuroprotection. Int. Rev. Neurobiol. 40: 233–258.

  22. 22.

    , , and 1996. Neuroprotection by aspirin and sodium salicylate through blockade of NF-kappa B activation. Science 274: 1383–1385.

  23. 23.

    1997. Development of the NMDA ion-channel blocker, aptiganel hydrochloride, as a neuroprotective agent for acute CNS injury. Int. Rev. Neurobiol. 40: 173–195.

  24. 24.

    , , , and 1996. Comparative patch-clamp studies with freshly dissociated rat hippocampal and striatal neurons on the NMDA receptor antagonistic effects of amantadine and memantine. Eur. J. Neurosci. 8: 446–454.

  25. 25.

    , and 1994. Memantine selectively depresses NMDA receptor-mediated responses of rat spinal neurones in vivo. Neurosci. Lett. 165: 37–40.

  26. 26.

    , , , , , et al. 1996. Chronic low-dose glutamate is toxic to retinal ganglion cells. Toxicity blocked by memantine. Invest. Ophthalmol. Vis. Sci. 37: 1618–1624.

  27. 27.

    , , and 1988. Glutamate antagonist therapy reduces neurologic deficits produced by focal central nervous system ischemia. Arch. Neurol. 45: 148–153.

  28. 28.

    , and 1980. Distribution and metabolism of the potential anti-Parkinson drug memantine in the human. J. Neural Transm. Suppl. 16: 143–148.

  29. 29.

    and 1997. Psychotogenicity and N-methyl-D-aspartate receptor antagonism: implications for neuroprotective pharmacotherapy. Biol. Psychiatry 41: 135–144.

  30. 30.

    and 1995. Cerebrospinal fluid and serum concentrations of the N-methyl-D-aspartate (NMDA) receptor antagonist memantine in man. Neurosci. Lett. 195: 137–139.

  31. 31.

    , , , , , et al. 1994. An all D-amino acid opioid peptide with central analgesic activity from a combinatorial library. Science 266: 2019–2022.

  32. 32.

    1985. General method for the rapid solid-phase synthesis of large numbers of peptides: specificity of antigen-antibody interaction at the level of individual amino acids. Proc. Natl. Acad. Sci. USA 82: 5131–5135.

  33. 33.

    , , and 1994. Peptide libraries: Determination of relative reaction rates of protected amino acids in competitive couplings. Biopolymers 34: 1681–1689.

  34. 34.

    , and 1983. SN2 deprotection of synthetic peptides with a low concentration of HF in dimethyl sulfide: evidence and application in peptide synthesis. Journal of the American Chemical Society 105: 6442–6455.

  35. 35.

    , , and 1986. Simplified procedure for carrying out simultaneous multiple hydrogen fluoride cleavages of protected peptide resins. Int. J. Pept. Protein Res. 27: 673–678.

  36. 36.

    and 1994. Structure-function relations in ligand-gated ion channels: Reconstitution in lipid bilayers and heterologous expression in Xenopus oocytes. Methods: A Companion to Methods in Enzymology 6: 60–69.

  37. 37.

    , and 1995. Molecular design of the N-methyl-D-aspartate receptor binding site for phencyclidine and dizolcipine. Proc. Natl. Acad. Sci. USA 92: 8021–8025.

  38. 38.

    , and 1996. A single tryptophan on M2 of glutamate receptor channels confers high permeability to divalent cations. Biophys. J. 71: 749–758.

  39. 39.

    , , , , , et al. 1992. Molecular cloning, chromosomal mapping, and functional expression of human brain glutamate receptors. Proc. Natl. Acad. Sci. USA 89: 1443–1447.

  40. 40.

    , , and 1993. Molecular cloning, functional expression, and pharmacological characterization of an N-methyl-D-aspartate receptor subunit from human brain. Proc. Natl. Acad. Sci. USA 90: 5057–5061.

  41. 41.

    , , , , , et al. 1994. Cloning, functional expression, and pharmacological characterisation of human cDNAs encoding NMDA receptor NR1 and NR2A subunits. J. Neurochem. 62: 2091–2098.

Download references

Author information

Author notes

    • Antonio V. Ferrer-Montiel
    •  & Jaime M. Merino

    These authors contributed equally to this work.

Affiliations

  1. Department of Biology, University of California, San Diego, La Jolla, CA 92093-0366.

    • Antonio V. Ferrer-Montiel
    • , Jaime M. Merino
    •  & Mauricio Montal
  2. Torrey Pines Institute for Molecular Studies, San Diego, CA 92121.

    • Sylvie E. Blondelle
    • , Enrique Perez-Paya
    •  & Richard A. Houghten
  3. Present address: Departament of Biochemistry and Molecular Biology, University of Valencia, 46100 Burjassot, Valencia, Spain.

    • Enrique Perez-Paya

Authors

  1. Search for Antonio V. Ferrer-Montiel in:

  2. Search for Jaime M. Merino in:

  3. Search for Sylvie E. Blondelle in:

  4. Search for Enrique Perez-Paya in:

  5. Search for Richard A. Houghten in:

  6. Search for Mauricio Montal in:

Corresponding author

Correspondence to Mauricio Montal.

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nbt0398-286

Further reading