Evidence for specific interactions between kavain and human cortical neurons monitored by fluorescence correlation spectroscopy
Evidence for specific interactions between kavain and human cortical neurons monitored by fluorescence correlation spectroscopy
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DOI：
10.1055/s-2000-11112
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发表时间：
2000-02-01
期刊：
影响因子：
2.7
通讯作者：
Häberlein, H
中科院分区：
文献类型：
--
作者：
Boonen, G;Pramanik, A;Häberlein, H
利用荧光相关光谱法首次发现了四甲基罗丹明标记的醉椒素衍生物与人皮层神经元之间的特异性相互作用。将在Nunc室中培养的人皮质神经元与1 nM染料标记的醉椒素衍生物一起孵育。孵育60分钟后，发现总结合为0.55 nM。在1 μ M非标记（+）-醉椒素存在下，总结合的50%被特异性置换。这些具体的相互作用的证据得到了验证的饱和实验。非线性Scatchard图和S形Hill图中的1.58 +/-0.07的n值均表明具有不同结合亲和力的结合位点。
Specific interactions between a tetramethylrhodamine-labeled kavain derivative and human cortical neurons were found for the first time using fluorescence correlation spectroscopy (FCS). Human cortical neurons, cultivated in Nunc(R) chambers, were incubated with 1 nM of the dye-labeled kavain derivative. A total binding of 0.55 nM was found after an incubation period of 60 min. 50% of the total binding was specifically displaced in the presence of 1 mu M non-labeled (+)-kavain. Evidence for these specific interactions was verified by a saturation experiment. Both the non-linear Scatchard plot and the n value of 1.58 +/- 0.07 in the sigmoid Hill plot indicate binding sites with different binding affinities.