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Immunological Detection of the NMDAR1 Glutamate Receptor Subunit Expressed in Human Embryonic Kidney 293 Cells and in Rat Brain
Author(s) -
Chazot Paul L.,
Cik Miroslav,
Stephenson F. Anne
Publication year - 1992
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1992.tb08364.x
Subject(s) - transfection , biology , receptor , microbiology and biotechnology , glutamate receptor , protein subunit , keyhole limpet hemocyanin , nmda receptor , embryonic stem cell , antibody , biochemistry , gene , immunology
The rat NMDAR1 ( N ‐methyl‐d‐aspartate receptor) was expressed transiently in human embryonic kidney cells. Transfected cell homogenates showed saturable [ 3 H]MK‐801 binding activity that was best fit by a single high‐affinity site with a K D of 9 n M and a 5 max of 113 fmol of binding sites/mg of protein. Antibodies raised against the peptide sequence NMDAR1 (929–938) coupled to keyhole limpet haemocyanin specifically recognised a single band with M r 117,000 in immunoblots from adult rat brain. In the transfected cells, the antibody recognised two bands: one with M r 117,000, which was coincident with that from brain membranes, and one with M r 97,000, which was identified as nonglycosylated NMDAR1 subunit. These results identify the NMDAR1 of rat brain and further show that the homooligomer binds MK‐801, albeit at low efficiency.