Novel bioluminescent binding assays for interaction studies of protein/peptide hormones with their receptors.

Liu, Ya-Li; Guo, Zhan-Yun

Protein/peptide hormones are the largest group of endogenous signaling molecules and exert various biological functions by binding to specific cell membrane receptors. To study the interactions between these hormones and their receptors, quantitative ligand-receptor binding assays have been widely used for decades. However, the assays conventionally relied on the use of radioligands, which have some major drawbacks and can only be used in laboratories with a radioactive material license. We recently developed novel bioluminescent binding assays for several protein/peptide hormones using the brightest bioluminescent reporter known to date, nanoluciferase (NanoLuc). The NanoLuc reporter can be either chemically conjugated to an appropriate position, or genetically fused at one terminus, of protein/peptide hormones. Compared to conventional radioligands, these bioluminescent ligands have higher sensitivity, better safety, and longer shelf lives, and thus, represent a novel class of non-radioactive tracers for quantitative receptor binding assays. In the present review, we provide some general considerations and specific examples for setting up the bioluminescent binding assays. Such techniques can be applied to other protein/peptide hormones in future to facilitate their interaction studies with their receptors.

BIOLUMINESCENCE; PROTEIN-protein interactions; PEPTIDE hormones; CELLULAR signal transduction; RECEPTOR-ligand complexes

Amino Acids, 2016, Vol 48, Issue 5, p1151

0939-4451

Academic Journal

10.1007/s00726-016-2220-5