Measurement of the millisecond activation switch of G protein-coupled receptors in living cells
Measurement of the millisecond activation switch of G protein-coupled receptors in living cells
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DOI：
10.1038/nbt838
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发表时间：
2003-07-01
影响因子：
46.9
通讯作者：
Lohse, MJ
中科院分区：
文献类型：
--
作者：
Vilardaga, JP;Bünemann, M;Lohse, MJ
激素和神经递质通过G蛋白偶联受体（GPCR）传递信号。尽管它们有共同的信号通路，但是它们引起的反应具有不同的时间模式。为了揭示这些差异的分子基础，我们开发了一种普遍适用的基于荧光的技术，用于实时监测活细胞中GPCR的激活开关。我们使用这种直接测量来研究α（2A）-肾上腺素能受体（α（2A）AR;神经递质）和甲状旁腺激素受体（PTHR;激素）的激活，并观察到比预期快得多的动力学：α（2A）AR类似于40 ms，PTHR类似于1 s。不同的开关时间与不同受体的生物学功能相一致。激动剂和拮抗剂可以迅速地打开或关闭受体，而部分激动剂只产生部分信号。这种方法允许在受体水平上比较激动剂和部分激动剂的固有活性，并提供GPCR毫秒激活时间的证据。
Hormones and neurotransmitters transduce signals through G protein-coupled receptors (GPCR). Despite their common signaling pathways, however, the responses they elicit have different temporal patterns. To reveal the molecular basis for these differences we have developed a generally applicable fluorescence-based technique for real-time monitoring of the activation switch of GPCRs in living cells. We used such direct measurements to investigate the activation of the alpha(2A)-adrenergic receptor (alpha(2A) AR; neurotransmitter) and the parathyroid hormone receptor (PTHR; hormone) and observed much faster kinetics than expected: similar to40 ms for the alpha(2A) AR and similar to1 s for the PTHR. The different switch times are in agreement with the different receptors' biological functions. Agonists and antagonists could rapidly switch the receptors on or off, whereas a partial agonist caused only a partial signal. This approach allows the comparison of agonist and partial agonist intrinsic activities at the receptor level and provides evidence for millisecond activation times of GPCRs.