1383430

Functional reconstitution of alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) receptors from rat brain.

Article date: 1992/11/1

PubMed ID: 1383430

Journal name: Journal of neurochemistry (ISSN: 0022-3042)

ABSTRACT

Glutamate receptors belonging to the subclass specifically activated by alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) were solubilized from rat forebrain membranes with Triton X-100 and partially purified through a series of three chromatographic steps. Specific [3H]AMPA binding increased 30-60-fold during the isolation procedure. A protein band recognized by antibodies against specific amino acid sequences of the glutamate receptor-A subunit was enriched with each purification step; the molecular mass of this band (105 kDa) corresponded to that of cloned AMPA receptor subunits. Photoaffinity labeling of forebrain membranes with 6-cyano-7-[3H]nitroquinoxaline-2,3-dione, a specific antagonist of the AMPA receptor, labeled a single band that comigrated with the immunolabeled protein. On reconstitution of the partially purified material into bilayer patches, single-channel current fluctuations were elicited by 300 nM AMPA and blocked by 1 microM 6,7-dinitroquinoxaline-2,3-dione.

Author List: Bahr B A, Vodyanoy V, Hall R A, Suppiramaniam V, Kessler M, Sumikawa K, Lynch G

Publication Types: Journal Article; Research Support, U.S. Gov't, Non-P.H.S.

Substances mentioned in the article: Lipid Bilayers; Receptors, AMPA; Receptors, Neurotransmitter; Ibotenic Acid; alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid;

Mesh terms: Animals; Ibotenic Acid/analogs & derivatives; Lipid Bilayers/chemistry; Prosencephalon/chemistry; Protein Binding; Rats; Receptors, AMPA; Receptors, Neurotransmitter/chemistry; alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid;

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