Membrane topology of the GluR1 glutamate receptor subunit: Epitope mapping by site-directed antipeptide antibodies

Elek Molnár, R. A Jeffrey McIlhinney, Agnès Baude, Z. Nusser, Péter Somogyi

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

In order to define the membrane topology of the GluR1 glutamate receptor subunit, we have examined the location of epitopes. Antibodies were produced against peptides corresponding to putative extracellular and intracellular segments of the rat brain GluR1 glutamate receptor subunit. Immunocytochemistry at the electron microscopic level in the dentate gyrus of the hippocampal formation showed that epitopes for the antiserum to the N- terminal part of the subunit are located at the extracellular face of the plasma membrane, whereas the antigenic determinants for the antiserum to the C-terminal part are found at the intracellular face of the postsynaptic membrane. Furthermore, antibodies to the N-terminal residues 253-267 reacted similarly with both intact and permeabilized synaptosomes, whereas the binding of antibodies to the C-terminal residues 877-889 increased about 1.6- fold following permeabilization. Our data suggest that the N- and C-terminal regions are located on the opposite side of the membrane and, therefore, the GluR1 subunit probably has an odd number of membrane spanning segments. The antibody cross-reactivities in different species and their effect on ligand binding activity were also established.

Original languageEnglish
Pages (from-to)683-693
Number of pages11
JournalJournal of Neurochemistry
Volume63
Issue number2
Publication statusPublished - Aug 1994

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Epitope Mapping
Glutamate Receptors
Epitopes
Topology
Membranes
Antibodies
Immune Sera
Synaptosomes
Dentate Gyrus
Cell membranes
Rats
Hippocampus
Brain
Immunohistochemistry
Cell Membrane
Electrons
Ligands
Peptides

Keywords

  • α-Amino-3-hydroxy-5-methyl-4-isoxazole propionate
  • Antipeptide antibodies
  • Dentategyrus
  • Hippocampus
  • Immunocytochemistry
  • Topology

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Membrane topology of the GluR1 glutamate receptor subunit : Epitope mapping by site-directed antipeptide antibodies. / Molnár, Elek; McIlhinney, R. A Jeffrey; Baude, Agnès; Nusser, Z.; Somogyi, Péter.

In: Journal of Neurochemistry, Vol. 63, No. 2, 08.1994, p. 683-693.

Research output: Contribution to journalArticle

Molnár, Elek ; McIlhinney, R. A Jeffrey ; Baude, Agnès ; Nusser, Z. ; Somogyi, Péter. / Membrane topology of the GluR1 glutamate receptor subunit : Epitope mapping by site-directed antipeptide antibodies. In: Journal of Neurochemistry. 1994 ; Vol. 63, No. 2. pp. 683-693.
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