Résumé
Seven-transmembrane domain (7TM) receptors have important functions in cell-cell communication and can assemble into dimers or oligomers. Such complexes may allow specific functional cross-talk through trans-activation of interacting 7TMs, but this hypothesis requires further validation. Herein, we used the GABA(B) receptor, which is composed of two distinct subunits, GABA(B1), which binds the agonist, and GABA(B2), which activates G proteins, as a model system. By using a novel orthogonal-labelling approach compatible with time-resolved FRET and based on ACP- and SNAP-tag technologies to verify the heterodimerization of wild-type and mutated GABAB subunits, we demonstrate the existence of a direct allosteric coupling between the 7TMs of GABA(B) heterodimers. Indeed, a GABA(B) receptor, in which the GABA(B2) extracellular domain was deleted, was still capable of activating G proteins. Furthermore, synthetic ligands for the GABA(B2) 7TM could increase agonist affinity at the GABA(B1) subunit in this mutated receptor. In addition to bringing new information on GABA(B) receptor activation, these data clearly demonstrate the existence of direct trans-activation between the 7TM of two interacting proteins. The EMBO Journal (2011) 30, 32-42. doi:10.1038/emboj.2010.270; Published online 9 November 2010