刘彦信, 李灵源, 金荫昌. 大鼠脑内阿片受体的溶脱及稀释对其结合活性的影响J. 药学学报, 1985, 20(10): 721-725.
引用本文: 刘彦信, 李灵源, 金荫昌. 大鼠脑内阿片受体的溶脱及稀释对其结合活性的影响J. 药学学报, 1985, 20(10): 721-725.
LIU Yan-Xin, LI Ling-Yuan, JIN Yin-Chang. SOLUBILIZATION OF OPIATE RECEPTOR AND THE EFFECT OF DILUTION ON BINDING ACTIVITYJ. Acta Pharmaceutica Sinica, 1985, 20(10): 721-725.
Citation: LIU Yan-Xin, LI Ling-Yuan, JIN Yin-Chang. SOLUBILIZATION OF OPIATE RECEPTOR AND THE EFFECT OF DILUTION ON BINDING ACTIVITYJ. Acta Pharmaceutica Sinica, 1985, 20(10): 721-725.

大鼠脑内阿片受体的溶脱及稀释对其结合活性的影响

SOLUBILIZATION OF OPIATE RECEPTOR AND THE EFFECT OF DILUTION ON BINDING ACTIVITY

  • 摘要: 用氚标记依托菲(3H-etor)或氚标记二苯羟乙酸喹咛酯(3H-QNB)与大鼠脑(去小脑)P2膜制备共同保温后,以Triton x-100溶脱,再用Sepharose 6 B层析,阿片受体及M胆碱受体洗脱峰位置相同,分子量皆约为450,000。以CHAPS溶脱P2制备,可获得有活性的阿片及M胆碱受体。溶脱液被Tris缓冲液(50 mM,pH 7.5)稀释后,阿片受体结合能力可提高25倍,如稀释液中含CHAPS(1 mM)结合能力变化不大。

     

    Abstract: Incubation of P2 membrane preparation of rat brain homogenenate followed by solubilization with Triton-x 100, centrifugation at 100,000g for 1 hr and chromatography by Sephadex-G 200 column of the supernatant led to an elution profile in which opiate receptors and muscarinic receptors were found in the same protein peak. The molecular weight of both receptors were estimated to he 450,000 daltons.Neither opiate nor muscarinic receptors could be obtained in the active from when Triton-x 100 was used as the solubilizing agent, but solubilization with CHAPS was successful. With CHAPS, the yield of opiate and muscarinic receptors were about 23% and 12% respectively. Dilution of the solubilized supernatant with Tris buffer (50 mM, pH 7.5) enhenced the binding capacity of opiate receptor about 25 folds, but when the dilution buffer contained CHAPS (1 mM), there was virtually no increase in binding acivity.

     

/

返回文章
返回