Functional reconstitution of alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) receptors from rat brain.
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.
Bahr
B A
BA
Center for the Neurobiology of Learning and Memory, University of California, Irvine 92717-3800.
Vodyanoy
V
V
Hall
R A
RA
Suppiramaniam
V
V
Kessler
M
M
Sumikawa
K
K
Lynch
G
G
D000818:Animals; D007051:Ibotenic Acid / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D008051:Lipid Bilayers / Q000737:chemistry; D016548:Prosencephalon / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D051381:Rats; D018091:Receptors, AMPA; D017981:Receptors, Neurotransmitter / Q000737:chemistry / Q000302:isolation & purification* / Q000502:physiology*; D018350:alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid
D008051:Lipid Bilayers; D018091:Receptors, AMPA; D017981:Receptors, Neurotransmitter; D007051:Ibotenic Acid; D018350:alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid