Brilliant Blue G selectively blocks ATP-gated rat P2X7 receptors
Brilliant Blue G selectively blocks ATP-gated rat P2X7 receptors
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DOI：
10.1124/mol.58.1.82
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发表时间：
2000-07-01
影响因子：
3.6
通讯作者：
Surprenant, A
中科院分区：
文献类型：
--
作者：
Jiang, LH;Mackenzie, AB;Surprenant, A
对几种P2 X受体的亚型具有选择性的拮抗剂很少，但需要这些拮抗剂来鉴定在天然细胞和组织上表达的受体。特别地，P2 X（4）和P2 X（7）受体亚单位共定位于免疫、上皮和外分泌腺细胞上，但两者对苏拉明和吡哆醛-5-磷酸-6-偶氮-2 '，4'-二磺酸衍生物相对不敏感。在这篇文章中，我们表明考马斯亮蓝G选择性抑制P2 X（7）受体与纳摩尔亲和力。我们在异源表达人或大鼠P2 X（1）、P2 X（2）、P2 X（3）、P2 X（2/3）、P2 X（4）、P2 X（1/5）和P2 X（7）受体的HEK 293细胞中测量了响应于P2 X受体激活的电流。亮蓝G对大鼠和人P2 X（7）受体产生非竞争性抑制，IC 50值分别为10和200 nM。抑制其他受体的IC 50值范围为2至>30 μ M;大鼠和人P2 X（4）受体的IC 50值分别为>10和3.2 μ M。考马斯亮蓝G还阻断了YO-PRO 1摄取和膜起泡，这与P2 X（7）受体的激活密切相关。因此，亮蓝G对大鼠P2 X（7）受体的效力至少是对大鼠P2 X（4）受体的1000倍。
There are few antagonists selective for subtypes of the several P2X receptors, but these are needed to identify the receptors expressed on native cells and tissues. In particular, P2X(4) and P2X(7) receptor subunits are colocalized on immune, epithelial, and exocrine gland cells, but both are relatively insensitive to suramin and pyridoxal-5-phosphate-6-azo-2',4'-disulfonic acid derivative. In this article, we show that Coomassie Brilliant Blue G selectively inhibits P2X(7) receptors with nanomolar affinity. We measured currents in response to P2X receptor activation in HEK293 cells heterologously expressing human or rat P2X(1), P2X(2), P2X(3), P2X(2/3), P2X(4), P2X(1/5), and P2X(7) receptors. Brilliant Blue G produced a noncompetitive inhibition of rat and human P2X(7) receptors with IC50 values of 10 and 200 nM, respectively. IC50 values for inhibition of the other receptors ranged from 2 to >30 mu M; the rat and human P2X(4) receptors showed IC50 values of >10 and 3.2 mu M. Coomassie Blue G also blocked YO-PRO1 uptake and membrane blebbing, which are uniquely associated with activation of P2X(7) receptors. Thus, Brilliant Blue G is at least 1000-fold more potent at rat P2X(7) receptors than at rat P2X(4) receptors.