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Protein‐protein interactions monitored in cells from transgenic mice using bioluminescence resonance energy transfer

Monitoring the dynamics of protein‐protein interactions in their natural environment remains a challenge. Resonance energy transfer approaches represent a promising avenue to directly probe these interactions in real time. The present study aims at establishing a proof of principle that bioluminescence resonance energy transfer (BRET) can be used to study the regulation of protein‐protein interaction in cells from transgenic animals. A transgenic mouse line coex‐pressing the β2‐adrenergic receptor fused to Renilla luciferase (β2AR‐Rluc) and βarrestin‐2 fused to a green fluorescent protein (GFP2‐βarr2) was generated. The fusion proteins were found to be functional in the transgenic animals and the β2AR‐Rluc maintained pharmacological properties, comparable to that of the native receptor. Sufficiently high luminescence signal was generated to allow detection of BRET in testis cells where the β2AR‐Rluc transgene was expressed at levels significantly higher than that of the endogenous receptor in this tissue but remain within physiological range when compared with other β2AR‐expressing tissues. Stimulation with a β‐adrenergic agonist led to a significant dose‐ and time‐dependent increase in BRET, which reflected ligand‐promoted recruitment of βarr2 to the receptor. Our study demonstrates that BRET can be used to monitor the dynamic regulation of protein‐protein interactions in cells derived from transgenic mice.—Audet, M., Lagacé, M., Silversides, D. W., Bouvier, M. Protein‐protein interactions monitored in cells from transgenic mice using bioluminescence resonance energy transfer. FASEB J. 24, 2829–2838 (2010). www.fasebj.org