Skip to main content
PMC home page
Biophysical Journal logoLink to Biophysical Journal
. 2002 Dec;83(6):3304–3314. doi: 10.1016/S0006-3495(02)75331-9

Staggering of subunits in NMDAR channels.

Alexander I Sobolevsky 1, LeeAnn Rooney 1, Lonnie P Wollmuth 1
PMCID: PMC1302406  PMID: 12496098

Abstract

Functional N-methyl-D-aspartate receptors (NMDARs) are heteromultimers formed by NR1 and NR2 subunits. The M3 segment, as contributed by NR1, forms the core of the extracellular vestibule, including binding sites for channel blockers, and represents a critical molecular link between ligand binding and channel opening. Taking advantage of the substituted cysteine accessibility method along with channel block and multivalent coordination, we studied the contribution of the M3 segment in NR2C to the extracellular vestibule. We find that the M3 segment in NR2C, like that in NR1, contributes to the core of the extracellular vestibule. However, the M3 segments from the two subunits are staggered relative to each other in the vertical axis of the channel. Compared to NR1, homologous positions in NR2C, including those in the highly conserved SYTANLAAF motif, are located about four amino acids more externally. The staggering of subunits may represent a key structural feature underlying the distinct functional properties of NMDARs.

Full Text

The Full Text of this article is available as a PDF (317.0 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Amar M., Perin-Dureau F., Neyton J. High-affinity Zn block in recombinant N-methyl-D-aspartate receptors with cysteine substitutions at the Q/R/N site. Biophys J. 2001 Jul;81(1):107–116. doi: 10.1016/S0006-3495(01)75684-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anson L. C., Chen P. E., Wyllie D. J., Colquhoun D., Schoepfer R. Identification of amino acid residues of the NR2A subunit that control glutamate potency in recombinant NR1/NR2A NMDA receptors. J Neurosci. 1998 Jan 15;18(2):581–589. doi: 10.1523/JNEUROSCI.18-02-00581.1998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Antonov S. M., Gmiro V. E., Johnson J. W. Binding sites for permeant ions in the channel of NMDA receptors and their effects on channel block. Nat Neurosci. 1998 Oct;1(6):451–461. doi: 10.1038/2167. [DOI] [PubMed] [Google Scholar]
  4. Ayalon G., Stern-Bach Y. Functional assembly of AMPA and kainate receptors is mediated by several discrete protein-protein interactions. Neuron. 2001 Jul 19;31(1):103–113. doi: 10.1016/s0896-6273(01)00333-6. [DOI] [PubMed] [Google Scholar]
  5. Beck C., Wollmuth L. P., Seeburg P. H., Sakmann B., Kuner T. NMDAR channel segments forming the extracellular vestibule inferred from the accessibility of substituted cysteines. Neuron. 1999 Mar;22(3):559–570. doi: 10.1016/s0896-6273(00)80710-2. [DOI] [PubMed] [Google Scholar]
  6. Benveniste M., Mayer M. L. Trapping of glutamate and glycine during open channel block of rat hippocampal neuron NMDA receptors by 9-aminoacridine. J Physiol. 1995 Mar 1;483(Pt 2):367–384. doi: 10.1113/jphysiol.1995.sp020591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Burnashev N., Schoepfer R., Monyer H., Ruppersberg J. P., Günther W., Seeburg P. H., Sakmann B. Control by asparagine residues of calcium permeability and magnesium blockade in the NMDA receptor. Science. 1992 Sep 4;257(5075):1415–1419. doi: 10.1126/science.1382314. [DOI] [PubMed] [Google Scholar]
  8. Chatterton Jon E., Awobuluyi Marc, Premkumar Louis S., Takahashi Hiroto, Talantova Maria, Shin Yeonsook, Cui Jiankun, Tu Shichun, Sevarino Kevin A., Nakanishi Nobuki. Excitatory glycine receptors containing the NR3 family of NMDA receptor subunits. Nature. 2002 Jan 30;415(6873):793–798. doi: 10.1038/nature715. [DOI] [PubMed] [Google Scholar]
  9. Chiamvimonvat N., Pérez-García M. T., Ranjan R., Marban E., Tomaselli G. F. Depth asymmetries of the pore-lining segments of the Na+ channel revealed by cysteine mutagenesis. Neuron. 1996 May;16(5):1037–1047. doi: 10.1016/s0896-6273(00)80127-0. [DOI] [PubMed] [Google Scholar]
  10. Dingledine R., Borges K., Bowie D., Traynelis S. F. The glutamate receptor ion channels. Pharmacol Rev. 1999 Mar;51(1):7–61. [PubMed] [Google Scholar]
  11. Doyle D. A., Morais Cabral J., Pfuetzner R. A., Kuo A., Gulbis J. M., Cohen S. L., Chait B. T., MacKinnon R. The structure of the potassium channel: molecular basis of K+ conduction and selectivity. Science. 1998 Apr 3;280(5360):69–77. doi: 10.1126/science.280.5360.69. [DOI] [PubMed] [Google Scholar]
  12. Hirai H., Kirsch J., Laube B., Betz H., Kuhse J. The glycine binding site of the N-methyl-D-aspartate receptor subunit NR1: identification of novel determinants of co-agonist potentiation in the extracellular M3-M4 loop region. Proc Natl Acad Sci U S A. 1996 Jun 11;93(12):6031–6036. doi: 10.1073/pnas.93.12.6031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hollmann M., Heinemann S. Cloned glutamate receptors. Annu Rev Neurosci. 1994;17:31–108. doi: 10.1146/annurev.ne.17.030194.000335. [DOI] [PubMed] [Google Scholar]
  14. Holm Richard H., Kennepohl Pierre, Solomon Edward I. Structural and Functional Aspects of Metal Sites in Biology. Chem Rev. 1996 Nov 7;96(7):2239–2314. doi: 10.1021/cr9500390. [DOI] [PubMed] [Google Scholar]
  15. Horenstein J., Wagner D. A., Czajkowski C., Akabas M. H. Protein mobility and GABA-induced conformational changes in GABA(A) receptor pore-lining M2 segment. Nat Neurosci. 2001 May;4(5):477–485. doi: 10.1038/87425. [DOI] [PubMed] [Google Scholar]
  16. Jones Kevin S., VanDongen Hendrika M. A., VanDongen Antonius M. J. The NMDA receptor M3 segment is a conserved transduction element coupling ligand binding to channel opening. J Neurosci. 2002 Mar 15;22(6):2044–2053. doi: 10.1523/JNEUROSCI.22-06-02044.2002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Karlin A., Akabas M. H. Substituted-cysteine accessibility method. Methods Enzymol. 1998;293:123–145. doi: 10.1016/s0076-6879(98)93011-7. [DOI] [PubMed] [Google Scholar]
  18. Karlin Arthur. Emerging structure of the nicotinic acetylcholine receptors. Nat Rev Neurosci. 2002 Feb;3(2):102–114. doi: 10.1038/nrn731. [DOI] [PubMed] [Google Scholar]
  19. Kashiwagi Keiko, Masuko Takashi, Nguyen Christopher D., Kuno Tomoko, Tanaka Ikuko, Igarashi Kazuei, Williams Keith. Channel blockers acting at N-methyl-D-aspartate receptors: differential effects of mutations in the vestibule and ion channel pore. Mol Pharmacol. 2002 Mar;61(3):533–545. doi: 10.1124/mol.61.3.533. [DOI] [PubMed] [Google Scholar]
  20. Kohda K., Wang Y., Yuzaki M. Mutation of a glutamate receptor motif reveals its role in gating and delta2 receptor channel properties. Nat Neurosci. 2000 Apr;3(4):315–322. doi: 10.1038/73877. [DOI] [PubMed] [Google Scholar]
  21. Krupp J. J., Vissel B., Heinemann S. F., Westbrook G. L. N-terminal domains in the NR2 subunit control desensitization of NMDA receptors. Neuron. 1998 Feb;20(2):317–327. doi: 10.1016/s0896-6273(00)80459-6. [DOI] [PubMed] [Google Scholar]
  22. Kuner T., Wollmuth L. P., Karlin A., Seeburg P. H., Sakmann B. Structure of the NMDA receptor channel M2 segment inferred from the accessibility of substituted cysteines. Neuron. 1996 Aug;17(2):343–352. doi: 10.1016/s0896-6273(00)80165-8. [DOI] [PubMed] [Google Scholar]
  23. Kuryatov A., Laube B., Betz H., Kuhse J. Mutational analysis of the glycine-binding site of the NMDA receptor: structural similarity with bacterial amino acid-binding proteins. Neuron. 1994 Jun;12(6):1291–1300. doi: 10.1016/0896-6273(94)90445-6. [DOI] [PubMed] [Google Scholar]
  24. Laube B., Hirai H., Sturgess M., Betz H., Kuhse J. Molecular determinants of agonist discrimination by NMDA receptor subunits: analysis of the glutamate binding site on the NR2B subunit. Neuron. 1997 Mar;18(3):493–503. doi: 10.1016/s0896-6273(00)81249-0. [DOI] [PubMed] [Google Scholar]
  25. Leszkiewicz D. N., Kandler K., Aizenman E. Enhancement of NMDA receptor-mediated currents by light in rat neurones in vitro. J Physiol. 2000 Apr 15;524(Pt 2):365–374. doi: 10.1111/j.1469-7793.2000.t01-1-00365.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Liu Y., Holmgren M., Jurman M. E., Yellen G. Gated access to the pore of a voltage-dependent K+ channel. Neuron. 1997 Jul;19(1):175–184. doi: 10.1016/s0896-6273(00)80357-8. [DOI] [PubMed] [Google Scholar]
  27. Liu Y., Jurman M. E., Yellen G. Dynamic rearrangement of the outer mouth of a K+ channel during gating. Neuron. 1996 Apr;16(4):859–867. doi: 10.1016/s0896-6273(00)80106-3. [DOI] [PubMed] [Google Scholar]
  28. Loring R. H., Dou Y. M., Lane W., Jones G. S., Jr, Stevenson K. J. Aromatic trivalent arsenicals: covalent yet reversible reagents for the agonist binding site of nicotinic receptors. Brain Res Mol Brain Res. 1992 Sep;15(1-2):113–120. doi: 10.1016/0169-328x(92)90158-8. [DOI] [PubMed] [Google Scholar]
  29. Mansour M., Nagarajan N., Nehring R. B., Clements J. D., Rosenmund C. Heteromeric AMPA receptors assemble with a preferred subunit stoichiometry and spatial arrangement. Neuron. 2001 Dec 6;32(5):841–853. doi: 10.1016/s0896-6273(01)00520-7. [DOI] [PubMed] [Google Scholar]
  30. Paoletti P., Ascher P. Mechanosensitivity of NMDA receptors in cultured mouse central neurons. Neuron. 1994 Sep;13(3):645–655. doi: 10.1016/0896-6273(94)90032-9. [DOI] [PubMed] [Google Scholar]
  31. Robert A., Irizarry S. N., Hughes T. E., Howe J. R. Subunit interactions and AMPA receptor desensitization. J Neurosci. 2001 Aug 1;21(15):5574–5586. doi: 10.1523/JNEUROSCI.21-15-05574.2001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Rosenmund C., Stern-Bach Y., Stevens C. F. The tetrameric structure of a glutamate receptor channel. Science. 1998 Jun 5;280(5369):1596–1599. doi: 10.1126/science.280.5369.1596. [DOI] [PubMed] [Google Scholar]
  33. Sobolevsky A. I., Koshelev S. G., Khodorov B. I. Probing of NMDA channels with fast blockers. J Neurosci. 1999 Dec 15;19(24):10611–10626. doi: 10.1523/JNEUROSCI.19-24-10611.1999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Sobolevsky A. I. Two-component blocking kinetics of open NMDA channels by organic cations. Biochim Biophys Acta. 1999 Jan 12;1416(1-2):69–91. doi: 10.1016/s0005-2736(98)00211-9. [DOI] [PubMed] [Google Scholar]
  35. Sobolevsky A., Koshelev S. Two blocking sites of amino-adamantane derivatives in open N-methyl-D-aspartate channels. Biophys J. 1998 Mar;74(3):1305–1319. doi: 10.1016/S0006-3495(98)77844-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Sobolevsky Alexander I., Beck Christine, Wollmuth Lonnie P. Molecular rearrangements of the extracellular vestibule in NMDAR channels during gating. Neuron. 2002 Jan 3;33(1):75–85. doi: 10.1016/s0896-6273(01)00560-8. [DOI] [PubMed] [Google Scholar]
  37. Solomon Edward I., Sundaram Uma M., Machonkin Timothy E. Multicopper Oxidases and Oxygenases. Chem Rev. 1996 Nov 7;96(7):2563–2606. doi: 10.1021/cr950046o. [DOI] [PubMed] [Google Scholar]
  38. Sprengel R., Aronoff R., Völkner M., Schmitt B., Mosbach R., Kuner T. Glutamate receptor channel signatures. Trends Pharmacol Sci. 2001 Jan;22(1):7–10. doi: 10.1016/s0165-6147(00)01588-1. [DOI] [PubMed] [Google Scholar]
  39. Subramaniam S., Donevan S. D., Rogawski M. A. Hydrophobic interactions of n-alkyl diamines with the N-methyl-D-aspartate receptor: voltage-dependent and -independent blocking sites. Mol Pharmacol. 1994 Jan;45(1):117–124. [PubMed] [Google Scholar]
  40. Villarroel A., Regalado M. P., Lerma J. Glycine-independent NMDA receptor desensitization: localization of structural determinants. Neuron. 1998 Feb;20(2):329–339. doi: 10.1016/s0896-6273(00)80460-2. [DOI] [PubMed] [Google Scholar]
  41. Vissel Bryce, Krupp Johannes J., Heinemann Stephen F., Westbrook Gary L. Intracellular domains of NR2 alter calcium-dependent inactivation of N-methyl-D-aspartate receptors. Mol Pharmacol. 2002 Mar;61(3):595–605. doi: 10.1124/mol.61.3.595. [DOI] [PubMed] [Google Scholar]
  42. Vlachová V., Zemková H., Vyklický L., Jr Copper modulation of NMDA responses in mouse and rat cultured hippocampal neurons. Eur J Neurosci. 1996 Nov;8(11):2257–2264. doi: 10.1111/j.1460-9568.1996.tb01189.x. [DOI] [PubMed] [Google Scholar]
  43. Wollmuth L. P., Kuner T., Sakmann B. Intracellular Mg2+ interacts with structural determinants of the narrow constriction contributed by the NR1-subunit in the NMDA receptor channel. J Physiol. 1998 Jan 1;506(Pt 1):33–52. doi: 10.1111/j.1469-7793.1998.00033.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Wollmuth L. P., Kuner T., Seeburg P. H., Sakmann B. Differential contribution of the NR1- and NR2A-subunits to the selectivity filter of recombinant NMDA receptor channels. J Physiol. 1996 Mar 15;491(Pt 3):779–797. doi: 10.1113/jphysiol.1996.sp021257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Yang J., Ellinor P. T., Sather W. A., Zhang J. F., Tsien R. W. Molecular determinants of Ca2+ selectivity and ion permeation in L-type Ca2+ channels. Nature. 1993 Nov 11;366(6451):158–161. doi: 10.1038/366158a0. [DOI] [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES