| First Author | Audet M | Year | 2010 |
| Journal | FASEB J | Volume | 24 |
| Issue | 8 | Pages | 2829-38 |
| PubMed ID | 20335229 | Mgi Jnum | J:162347 |
| Mgi Id | MGI:4818735 | Doi | 10.1096/fj.09-144816 |
| Citation | Audet M, et al. (2010) Protein-protein interactions monitored in cells from transgenic mice using bioluminescence resonance energy transfer. FASEB J 24(8):2829-38 |
| abstractText | 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 coexpressing the beta(2)-adrenergic receptor fused to Renilla luciferase (beta(2)AR-Rluc) and betaarrestin-2 fused to a green fluorescent protein (GFP2-betaarr2) was generated. The fusion proteins were found to be functional in the transgenic animals and the beta(2)AR-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 beta(2)AR-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 beta(2)AR-expressing tissues. Stimulation with a beta-adrenergic agonist led to a significant dose- and time-dependent increase in BRET, which reflected ligand-promoted recruitment of betaarr2 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., Lagace, M., Silversides, D. W., Bouvier, M. Protein-protein interactions monitored in cells from transgenic mice using bioluminescence resonance energy transfer. |
