Solubilization and characterization of a [3H]hemicholinium-3 binding site in rat brain.
Solubilization and characterization of a [3H]hemicholinium-3 binding site in rat brain.
复制标题
大鼠脑中 [3H]hemicholinium-3 结合位点的溶解和表征。
DOI：
10.1111/j.1471-4159.1988.tb02475.x
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发表时间：
1988
影响因子：
4.7
通讯作者：
Coyle,JT
中科院分区：
文献类型：
--
作者：
Yamada,K;Saltarelli,MD;Coyle,JT
钠依赖性高亲和力[3 H]-hemicholinium-3（[3 H]HCH-3）结合位点通过0.2%脱氧胆酸盐从大鼠纹状体突触质膜中溶解。脱氧胆酸盐对[3 H]HCH-3结合位点的增溶作用取决于去污剂浓度和增溶介质的离子强度。[3 H]HCH-3与溶解制剂的特异性结合是钠和氯依赖性和饱和性的，表现出14.2 nM的亲和力和695 fmol/mg蛋白质的容量（Bmax）。胆碱和其他类似物以浓度依赖性方式抑制特异性[3 H]HCh-3与溶解制剂的结合，在粗突触膜中观察到类似的效力等级顺序。已知破坏蛋白质和脂质部分的处理会导致特异性[3 H]HCH-3结合减少。这些结果表明，溶解的[3 H]HCh-3结合位点的特征与膜结合位点的特征相似。
A sodium‐dependent high‐affinity [3H]‐hemicholinium‐3 ([3H]HCh‐3) binding site was solubilized from rat striatal synaptic plasma membranes by 0.2% deoxycholate. Deoxycholate solubilization of the [3H]HCh‐3 binding site was dependent upon both detergent concentration and ionic strength of the solubilization medium. Specific [3H]HCh‐3 binding to the solubilized preparation was both sodium‐ and chloride‐dependent and saturable, exhibiting an affinity of 14.2 nMand a capacity (Bmax) of 695 fmol/mg protein. Choline and other analogs inhibited specific [3H]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 [3H]HCh‐3 binding. These results suggest that the characteristics of the solubilized [3H]HCh‐3 binding site are similar to those of the membrane‐bound site.