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通过放射性标记的GTP结合测量G蛋白偶联的受体信号
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
Measuring G-protein-coupled Receptor Signaling via Radio-labeled GTP Binding
DOI:

10:13 min

June 09, 2017

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Chapters

  • 00:05Title
  • 00:37Expression of Recombinant HA-MOR1, Cell Fractionation, and Membrane Collection
  • 03:55[35S]GTPγS Binding
  • 06:05Membrane Filtration
  • 06:59Liquid Scintillation Counting
  • 07:57Results: Characterization of G-protein-coupled Receptors by Monitoring Radio-labled GTP Binding
  • 09:01Conclusion

Summary

Automatic Translation

三磷酸鸟苷(GTP)结合是G蛋白偶联受体(GPCR)激活中最早的事件之一。该方案描述了如何通过监测放射性标记的GTP类似物[ 35 S]鸟苷-5'-O-(3-硫代)三磷酸([ 35 S]GTPγS)的结合在药理学上表征特异性GPCR-配体相互作用对感兴趣配体的反应。

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