Background: Receptor heteromerisation is the phenomenon whereby two different receptors form a functional complex that attains unique pharmacological properties. Consequently, receptor heteromers can be considered novel drug targets, with the potential to achieve greater therapeutic selectivity and specificity. The angiotensin II type 2 (AT2) receptor and bradykinin type 2 (B2) receptor are both promising drug targets in their own right (for cardiovascular and other diseases), and the existence of a heteromer that forms between the two could allow improved specificity of targeting. The two receptors have some overlapping pharmacology, such as nitric oxide-mediated vasodilation, and there is some evidence that observed functional interactions may occur as a result of heteromerization (1).
Aim: Investigation of evidence for heteromerization of the AT2 receptor and the B2 receptor.
Methods: AT2 receptor and B2 receptor pharmacology and potential heteromerisation was investigated in HEK293FT cells using various bioluminescence resonance energy transfer (BRET)-proximity based assays. These assays allow real time, live cell monitoring of proximity between biomolecules of interest, such as receptors and signalling proteins, or receptors and ligands, and therefore produce physiologically relevant, real time pharmacological data.
Results: Heteromerisation of the AT2 receptor and B2 receptor was confirmed with assays showing recruitment of various signalling and regulatory proteins proximal to AT2 receptors only upon B2 receptor coexpression and activation. Additionally, the close proximity between the two receptors was also demonstrated with the NanoBRET ligand-binding assay (2). Attainment of novel pharmacology (Gaz coupling) was observed upon heteromerisation, which did not occur with either receptor alone.
Discussion: Our study has confirmed the heteromerisation of the AT2 receptor and B2 receptor, and showed that this results in the attainment of novel pharmacology. Further studies will aim to reveal the physiological consequences of this heteromerisation.