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Solubilization and Characterization of a [ 3 H]Hemicholinium‐3 Binding Site in Rat Brain
Author(s) -
Yamada Kiyofumi,
Saltarelli Mario D.,
Coyle Joseph T.
Publication year - 1988
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.1471-4159.1988.tb02475.x
Subject(s) - solubilization , binding site , chemistry , membrane , sodium , choline , biochemistry , chaps , synaptic membrane , ionic strength , chromatography , biophysics , biology , aqueous solution , organic chemistry
A sodium‐dependent high‐affinity [ 3 H]‐hemicholinium‐3 ([ 3 H]HCh‐3) binding site was solubilized from rat striatal synaptic plasma membranes by 0.2% deoxycholate. Deoxycholate solubilization of the [ 3 H]HCh‐3 binding site was dependent upon both detergent concentration and ionic strength of the solubilization medium. Specific [ 3 H]HCh‐3 binding to the solubilized preparation was both sodium‐ and chloride‐dependent and saturable, exhibiting an affinity of 14.2 n M and a capacity ( B max ) of 695 fmol/mg protein. Choline and other analogs inhibited specific [ 3 H]HCh‐3 binding to the solubilized preparation in a concentration‐dependent manner with the similar rank order of potency observed in crude synaptic membranes. Treatments known to disrupt both protein and lipid moieties resulted in diminished specific [ 3 H]HCh‐3 binding. These results suggest that the characteristics of the solubilized [ 3 H]HCh‐3 binding site are similar to those of the membrane‐bound site.