Adenosine plays important roles in the pain signal transduction by activating adenosine receptors of two subtypes of A1 and A2A. In this study, FRET system based independent emission -spectral spectral spectral unmixing method (Iem-spFRET) was set up and used to measure the energy transfer from A1R to A2AR. The energy transfer efficiency calculated by Iem-spFRET is about 17.44%. All the above date and results demonstrate that FRET with special designed fluorescence proteins could be used to investigate the interaction between adenosine receptors.
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