用[3H]酪氨酸-D-丙氨酸-甘氨酸-（甲基）苯丙氨酸-甘醇（DAMGO）的氯甲基酮衍生物进行阿片样物质受体标记II：3H-酪氨酸-D-丙氨酸-甘氨酸-（甲基）苯丙氨酸氯甲基酮对μ阿片样物质结合位点的共价标记
Opioid receptor labeling with the chloromethyl ketone derivative of [3H]Tyr-D-Ala-Gly-(Me)Phe-Gly-ol (DAMGO) II: Covalent labeling of mu opioid binding site by 3H-Tyr-D-Ala-Gly-(Me)Phe chloromethyl ketone.
作者信息
Oktem H A, Moitra J, Benyhe S, Toth G, Lajtha A, Borsodi A
机构信息
Institute of Biochemistry, Hungarian Academy of Sciences, Szeged.
出版信息
Life Sci. 1991;48(18):1763-8. doi: 10.1016/0024-3205(91)90214-v.
Opioid receptors of rat brain membranes were prelabeled with 3H-Tyr-D-Ala2-(Phe4)-Gly-CH2Cl, a chloromethyl ketone derivative of enkephalin, and solubilized in 1% digitonin. Hydrodynamic parameters of the receptor detergent complex derived from gel filtration and sucrose density gradient ultracentrifugation were found to be 51 A and 8.7 S, respectively, and the size was estimated to be about 200 kDa. Sodium dodecyl sulfate gel electrophoresis followed by fluorography revealed specific alkylation of a major protein at 58 kDa.
大鼠脑膜的阿片受体先用脑啡肽的氯甲基酮衍生物3H-Tyr-D-Ala2-(Phe4)-Gly-CH2Cl进行预标记，然后溶解于1%的洋地黄皂苷中。通过凝胶过滤和蔗糖密度梯度超速离心得到的受体去污剂复合物的流体动力学参数分别为51 Å和8.7 S，估计其大小约为200 kDa。十二烷基硫酸钠凝胶电泳后进行荧光自显影，结果显示主要蛋白质在58 kDa处发生特异性烷基化。
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