Solubilization of the atrial muscarinic acetylcholine receptor: a new detergent system and rapid assays.
Solubilization of the atrial muscarinic acetylcholine receptor: a new detergent system and rapid assays.
复制标题
心房毒蕈碱乙酰胆碱受体的溶解：新的去污剂系统和快速测定。
DOI：
10.1016/0003-2697(81)90014-2
复制
发表时间：
1981
影响因子：
2.9
通讯作者：
Schimerlik,MI
中科院分区：
文献类型：
--
作者：
Cremo,CR;Herron,GS;Schimerlik,MI
毒蕈碱乙酰胆碱受体，确定氚化的L-奎宁环二苯乙酸（1-[3 H]QNB）结合，溶解从猪心房膜使用5：1（w/w）的比例毛地黄皂苷和胆酸盐。溶解的受体溶液的特异性结合活性通常超过每克蛋白质1.0 nmol l-[3 H]QNB位点，代表75-98%的总位点回收率和未经处理的心房膜的2 - 3倍富集。设计了两种快速测定洗涤剂提取物结合活性的方法，并与平衡透析法进行了比较。所有三种方法都得到了类似的结果。根据方法和温度，通过三种方法测定的1-[3 H]QNB的溶解受体的平衡解离常数从230 μ m到450 μ m不等。
The muscarinic acetylcholine receptor, identified by tritiated l-quinuclidinyl benzilate (l-[3H]QNB) binding, was solubilized from porcine atrial membranes using a 5:1 (w w ) ratio of digitonin and cholate. Specific binding activities of the solubilized receptor solutions usually exceeded 1.0 nmol l-[3H]QNB sites per gram of protein, representing 75–98% total site recovery and a two- to threefold enrichment over untreated atrial membranes. Two rapid assays for measuring the binding activities of detergent extracts were devised and compared with equilibrium dialysis. All three methods gave similar results. The equilibrium dissociation constant of the solubilized receptor for l-[3H]QNB as determined by the three methods varied from 230 to 450 pm depending on the method and temperature.