Influence of fluorophore and linker composition on the pharmacology of fluorescent adenosine A1 receptor ligands.
Influence of fluorophore and linker composition on the pharmacology of fluorescent adenosine A1 receptor ligands.
复制标题
DOI：
10.1111/j.1476-5381.2009.00488.x
复制
发表时间：
2010-02
影响因子：
7.3
通讯作者：
Hill SJ
中科院分区：
文献类型：
--
作者：
Baker JG;Middleton R;Adams L;May LT;Briddon SJ;Kellam B;Hill SJ
基于荧光的技术的引入，特别是荧光配体的开发，使得在单细胞和单分子水平上研究G蛋白偶联受体药理学成为可能。本研究评估了接头和荧光团的理化性质如何影响荧光激动剂和拮抗剂的药理学特性。使用稳定表达人腺苷 A1 受体和环状 3',5' 腺苷单磷酸反应元件分泌的胎盘碱性磷酸酶 (CRE-SPAP) 报告基因的中国仓鼠卵巢细胞，以及全细胞 [3H]-8-环戊基-1,3-二丙基黄嘌呤 (DPCPX) 放射性配体结合，评估一系列基于拮抗剂黄嘌呤的荧光配体的药理学特性胺同源物 (XAC) 和激动剂 5'（N-乙基甲酰胺基）腺苷 (NECA)。具有不同荧光团但接头长度相同的 NECA 和 XAC 衍生物与腺苷 A1 受体的结合特性显示出显着差异。 BODIPY 630/650 衍生物具有最高的亲和力。连接体长度也会影响药理学特性，具体取决于所使用的荧光团。特别是在荧光激动剂中，分别使用丹磺酰和 BODIPY 630/650 衍生物的大或小接头可以实现更高的激动剂效力。荧光配体的药理学受到荧光团和相关接头的严重影响。此外，我们的数据强烈表明，荧光团/连接体配对的物理化学特性决定了荧光团在目标受体环境中的位置，并且这与所得荧光的环境敏感性一起最终可能决定其作为荧光探针的实用性。本文是药理学成像主题部分的一部分。要查看该主题部分的社论，请访问 http://dx.doi.org/10.1111/j.1476-5381.2010.00685.x
The introduction of fluorescence-based techniques, and in particular the development of fluorescent ligands, has allowed the study of G protein-coupled receptor pharmacology at the single cell and single molecule level. This study evaluated how the physicochemical nature of the linker and the fluorophore affected the pharmacological properties of fluorescent agonists and antagonists. Chinese hamster ovary cells stably expressing the human adenosine A1 receptor and a cyclic 3′,5′ adenosine monophosphate response element-secreted placental alkaline phosphatase (CRE-SPAP) reporter gene, together with whole cell [3H]-8-cyclopentyl-1,3-dipropylxanthine (DPCPX) radioligand binding, were used to evaluate the pharmacological properties of a range of fluorescent ligands based on the antagonist xanthine amine congener (XAC) and the agonist 5′ (N-ethylcarboxamido) adenosine (NECA). Derivatives of NECA and XAC with different fluorophores, but equivalent linker length, showed significant differences in their binding properties to the adenosine A1 receptor. The BODIPY 630/650 derivatives had the highest affinity. Linker length also affected the pharmacological properties, depending on the fluorophore used. Particularly in fluorescent agonists, higher agonist potency could be achieved with large or small linkers for dansyl and BODIPY 630/650 derivatives, respectively. The pharmacology of a fluorescent ligand was critically influenced by both the fluorophore and the associated linker. Furthermore, our data strongly suggest that the physicochemical properties of the fluorophore/linker pairing determine where in the environment of the target receptor the fluorophore is placed, and this, together with the environmental sensitivity of the resulting fluorescence, may finally decide its utility as a fluorescent probe. This article is part of a themed section on Imaging in Pharmacology. To view the editorial for this themed section visit http://dx.doi.org/10.1111/j.1476-5381.2010.00685.x
登录
查看更多内容
DOI：
10.1016/0006-291x(92)91285-x
发表时间：
1992-09-16
影响因子：
3.1
作者：
LIBERT, F;VANSANDE, J;MENDLA, KD
通讯作者：
MENDLA, KD
影响因子：
4.8
作者：
Harikumar, KG;Pinon, DL;Miller, LJ
通讯作者：
Miller, LJ
DOI：
10.1073/pnas.0400420101
发表时间：
2004-03-30
影响因子：
11.1
作者：
Briddon, SJ;Middleton, RJ;Hill, SJ
通讯作者：
Hill, SJ
影响因子：
7.3
作者：
Cordeaux, Y;IJzerman, AP;Hill, SJ
通讯作者：
Hill, SJ
影响因子：
7.3
作者：
Middleton, Richard J.;Briddon, Stephen J.;Kellam, Barrie
通讯作者：
Kellam, Barrie