Molecular Endocrinology and Pharmacology Group, Harry Perkins Institute of Medical Research, Nedlands, WA 6009, Australia.
Centre for Medical Research, The University of Western Australia, Crawley, WA 6009, Australia.
Int J Mol Sci. 2021 Jan 22;22(3):1082. doi: 10.3390/ijms22031082.
Receptor heteromerization is the formation of a complex involving at least two different receptors with pharmacology that is distinct from that exhibited by its constituent receptor units. Detection of these complexes and monitoring their pharmacology is crucial for understanding how receptors function. The Receptor-Heteromer Investigation Technology (Receptor-HIT) utilizes ligand-dependent modulation of interactions between receptors and specific biomolecules for the detection and profiling of heteromer complexes. Previously, the interacting biomolecules used in Receptor-HIT assays have been intracellular proteins, however in this study we have for the first time used bioluminescence resonance energy transfer (BRET) with fluorescently-labeled ligands to investigate heteromerization of receptors on the cell surface. Using the Receptor-HIT ligand binding assay with NanoBRET, we have successfully investigated heteromers between the angiotensin II type 1 (AT) receptor and the β adrenergic receptor (AT-βAR heteromer), as well as between the AT and angiotensin II type 2 receptor (AT-AT heteromer).
受体异质二聚化是指至少两种具有不同药理学特性的受体形成复合物的过程，其药理学特性与组成受体单元的药理学特性不同。检测这些复合物并监测其药理学特性对于理解受体的功能至关重要。受体异质二聚化研究技术（Receptor-HIT）利用配体依赖性调节受体与特定生物分子之间的相互作用，用于检测和分析异质二聚体复合物。以前，Receptor-HIT 测定中使用的相互作用生物分子是细胞内蛋白，但在这项研究中，我们首次使用带有荧光标记配体的生物发光共振能量转移（BRET）来研究细胞表面受体的异质二聚化。我们使用 NanoBRET 进行 Receptor-HIT 配体结合测定，成功地研究了血管紧张素 II 型 1（AT）受体和β肾上腺素能受体（AT-βAR 异质二聚体）之间以及 AT 和血管紧张素 II 型 2 受体（AT-AT 异质二聚体）之间的异质二聚体。
