Reassessment of [3H]glutamate binding to human brain membrane preparations.
We studied [3H]glutamate binding to human brain membrane preparations. We found that the binding assay of [3H]glutamate should be done at 4 degrees C using glass-fiber filters pretreated with polyethyleneimine, sterile incubation buffer, and membrane preparations pretreated with a detergent. Under these assay conditions, we have characterized the three subtypes of glutamate receptors: N-methyl-D-aspartate-, quisqualate- and kainate-sensitive binding sites. Scatchard analysis revealed that the N-methyl-D-aspartate-sensitive [3H]glutamate binding sites consisted of a single component with a dissociation constant (Kd) of 28.2 +/- 2.8 nM and total concentration (Bmax) of 36.2 +/- 3.6 fmol/mg protein (N = 8). These data were much different from those presented in previous studies.
Department of Neurobiology, Kyoto Pharmaceutical University, Japan.
R
R
Fukunaga
H
H
Ninomiya
S
S
Shimohama
J
J
Kimura
M
M
Kameyama
T
T
Taniguchi
D000368:Aged; D001921:Brain / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D005971:Glutamates / Q000378:metabolism*; D018698:Glutamic Acid; D006801:Humans; D007608:Kainic Acid / Q000378:metabolism; D007700:Kinetics; D016318:Quisqualic Acid / Q000378:metabolism; D018091:Receptors, AMPA; D017470:Receptors, Glutamate; D018092:Kainic Acid Receptors; D016194:Receptors, N-Methyl-D-Aspartate / Q000378:metabolism; D017981:Receptors, Neurotransmitter / Q000378:metabolism*
D005971:Glutamates; D018698:Glutamic Acid; D007608:Kainic Acid; D016318:Quisqualic Acid; D018091:Receptors, AMPA; D017470:Receptors, Glutamate; D018092:Kainic Acid Receptors; D016194:Receptors, N-Methyl-D-Aspartate; D017981:Receptors, Neurotransmitter