Direct radiolabeling by [3H]quisqualic acid of group I metabotropic glutamate receptor in rat brain synaptic membranes.
[3H]Quisqualic acid (QA) was synthesized and used to label metabotropic glutamate receptor (mGluR) in rat brain synaptic membranes in the presence of three different ionotropic glutamate receptor agonists at respective saturating concentrations. Of several mGluR agonists tested, group I agonists were more potent in displacing [3H]QA binding than group II and group III agonists in the presence of the three ionotropic agonists. [3H]QA binding was markedly inhibited by guanine nucleotide analogues in a concentration-dependent manner at a concentration range of 10 nM to 1 mM. Scatchard analysis revealed that [3H]QA binding consisted of a single component with a K(d) of 50.9+/-5.3 nM and a B(max) of 431. 6+/-18.3 fmol/mg protein. These results suggest that [3H]QA indeed labels group I mGluR functionally coupled to GTP binding protein in rat brain synaptic membranes when determined under the experimental conditions employed.
Hinoi
E
E
Department of Molecular Pharmacology, Kanazawa University Faculty of Pharmaceutical Sciences, 13-1 Takara-machi, Kanazawa, 920-0934, Ishikawa, Japan.
Ogita
K
K
Takeuchi
Y
Y
Ohashi
H
H
Maruyama
T
T
Yoneda
Y
Y
D000818:Animals; D001921:Brain / Q000378:metabolism*; D018690:Excitatory Amino Acid Agonists / Q000378:metabolism*; D006160:Guanosine Triphosphate / Q000378:metabolism; D008297:Male; D016318:Quisqualic Acid / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D018094:Receptors, Metabotropic Glutamate / Q000378:metabolism*; D013570:Synaptic Membranes / Q000378:metabolism*
D018690:Excitatory Amino Acid Agonists; D018094:Receptors, Metabotropic Glutamate; C104077:metabotropic glutamate receptor type 1; D006160:Guanosine Triphosphate; D016318:Quisqualic Acid