Rinken A A, Langel Iu L, Iarv Ia L
Biokhimiia. 1987 Feb;52(2):303-10.
The possibilities to solubilize the rat brain cortex muscarinic acetylcholine receptor and its complex with [3H]-L-quinuclidinyl benzilate (QNB) were studied, using 14 detergents. It was shown that the native muscarinic cholinoreceptor was solubilized in addition to digitonin, also by CHAPS, with a 6% yield. Besides, the receptor-QNB complex was solubilized with the detergents Triton X-100, -102, -114, -165 (with 30% and 50% yields) and within a narrow concentration range with sodium dodecyl sulfate (50% yield). Some detergents of the Tween series, e.g., Triton X-45 and -305, as well as sodium deoxycholate and sodium oxycholate, did not solubilize the native receptor and its complex with QNB. It was found that yield of receptor solubilization did not exceed half of the total number of the receptor sites in the membranes, despite the fact that different concentrations of detergents were applied. The solubilization yield did not increase, when different mixtures of detergents were used. It was assumed that incomplete solubilization of the receptor protein reflects its heterogeneity in the membrane structure.
使用14种去污剂研究了溶解大鼠脑皮质毒蕈碱型乙酰胆碱受体及其与[3H]-L-奎宁环基苯甲酸酯(QNB)复合物的可能性。结果表明,除了洋地黄皂苷外,CHAPS也能溶解天然毒蕈碱型胆碱受体,产率为6%。此外,受体-QNB复合物可用去污剂Triton X-100、-102、-114、-165溶解(产率分别为30%和50%),在较窄的浓度范围内,十二烷基硫酸钠也可溶解(产率为50%)。吐温系列的一些去污剂,如Triton X-45和-305,以及脱氧胆酸钠和牛磺胆酸钠,不能溶解天然受体及其与QNB的复合物。结果发现,尽管使用了不同浓度的去污剂,但受体溶解产率不超过膜中受体位点总数的一半。当使用不同的去污剂混合物时,溶解产率并未增加。据推测,受体蛋白的不完全溶解反映了其在膜结构中的异质性。