New fluorescent adenosine A1-receptor agonists that allow quantification of ligand-receptor interactions in microdomains of single living cells
New fluorescent adenosine A1-receptor agonists that allow quantification of ligand-receptor interactions in microdomains of single living cells
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DOI：
10.1021/jm061279i
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发表时间：
2007-02-22
影响因子：
7.3
通讯作者：
Kellam, Barrie
中科院分区：
文献类型：
--
作者：
Middleton, Richard J.;Briddon, Stephen J.;Kellam, Barrie
荧光光谱学正在成为一个有价值的除了阵列的技术可用于审查配体-受体相互作用的生物系统。特别是，扫描共聚焦显微镜和荧光相关光谱（FCS）允许非侵入性成像和量化这些相互作用在单个活细胞。为了满足对荧光标记配体的新兴需求以支持这些技术，我们开发了一系列针对人类腺苷A（1）受体的红色发射激动剂，这些激动剂总体上是腺苷或腺苷的N-6-氨基烷基衍生物5 '-N-乙基甲酰胺。结合市售荧光团BODIPY [630/650]的激动剂保留了强效和有效的激动剂活性，如通过其抑制表达人腺苷A（1）受体的中国仓鼠卵巢细胞中cAMP蓄积的能力所证明的。使用共聚焦显微镜完成了在细胞膜上的配体-受体相互作用的可视化和确认，并且通过定量细胞膜小区域中的激动剂结合来证明它们在FCS中使用的适用性。
Fluorescence spectroscopy is becoming a valuable addition to the array of techniques available for scrutinizing ligand-receptor interactions in biological systems. In particular, scanning confocal microscopy and fluorescence correlation spectroscopy (FCS) allow the noninvasive imaging and quantification of these interactions in single living cells. To address the emerging need for fluorescently labeled ligands to support these technologies, we have developed a series of red-emitting agonists for the human adenosine A(1)-receptor that, collectively, are N-6-aminoalkyl derivatives of adenosine or adenosine 5'-N-ethyl carboxamide. The agonists, which incorporate the commercially available fluorophore BODIPY [630/650], retain potent and efficacious agonist activity, as demonstrated by their ability to inhibit cAMP accumulation in chinese hamster ovary cells expressing the human adenosine A(1)-receptor. Visualization and confirmation of ligand-receptor interactions at the cell membrane were accomplished using confocal microscopy, and their suitability for use in FCS was demonstrated by quantification of agonist binding in small areas of cell membrane.