title: NMDAR channel segments forming the extracellular vestibule inferred from the accessibility of substituted cysteines.
abstract: In NMDA receptor channels, the M2 loop forms the narrow constriction and the cytoplasmic vestibule. The identity of an extracellular vestibule leading toward the constriction remained unresolved. Using the substituted cysteine accessibility method (SCAM), we identified channel-lining residues of the NR1 subunit in the region preceding M1 (preM1), the C-terminal part of M3 (M3C), and the N-terminal part of M4 (M4N). These residues are located on the extracellular side of the constriction and, with one exception, are exposed to the pore independently of channel activation, suggesting that the gate is at the constriction or further cytoplasmic to it. Permeation of Ca2+ ions was decreased by mutations in M3C and M4N, but not by mutations in preM1, suggesting a functionally distinct contribution of the segments to the extracellular vestibule of the NMDA receptor channel.
authors: Abteilung Molekulare Neurobiologie, Max-Planck-Institut für medizinische Forschung, Heidelbeg, Germany.
authors: C
authors: C
authors: Beck
authors: L P
authors: LP
authors: Wollmuth
authors: P H
authors: PH
authors: Seeburg
authors: B
authors: B
authors: Sakmann
authors: T
authors: T
authors: Kuner
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002463:Cell Membrane Permeability / Q000502:physiology; D003545:Cysteine / Q000737:chemistry / Q000235:genetics*; D003593:Cytoplasm / Q000235:genetics / Q000502:physiology; D005110:Extracellular Space / Q000502:physiology*; D018698:Glutamic Acid / Q000494:pharmacology; D007202:Indicators and Reagents / Q000737:chemistry; D008564:Membrane Potentials / Q000502:physiology; D008698:Mesylates / Q000737:chemistry; D008969:Molecular Sequence Data; D009154:Mutation; D009865:Oocytes; D018408:Patch-Clamp Techniques; D016194:Receptors, N-Methyl-D-Aspartate / Q000235:genetics / Q000502:physiology*; D014981:Xenopus
chemical_list: D007202:Indicators and Reagents; D008698:Mesylates; D016194:Receptors, N-Methyl-D-Aspartate; D018698:Glutamic Acid; D003545:Cysteine; D002118:Calcium