Bioluminescent Ligand–Receptor Binding Assays for Protein or Peptide Hormones
Bioluminescence has been widely used in biomedical research due to its high sensitivity, low background, and broad linear range. In recent studies, we applied bioluminescence to ligand–receptor binding assays for some protein or peptide hormones based on a newly developed small monomeric Nanoluciferase (NanoLuc) reporter that has the so far brightest bioluminescence. The conventional ligand–receptor binding assays rely on radioligands that have drawbacks, such as radioactive hazards and short shelf lives. In contrast, the novel bioluminescent binding assays use the NanoLuc-based protein or peptide tracers that are safe, stable, and ultrasensitive. Thus, the novel bioluminescent ligand–receptor binding assay would be applied to more and more protein or peptide hormones for ligand–receptor interaction studies in future. In the present article, we provided detailed protocols for setting up the novel bioluminescent ligand–receptor binding assays using two representative protein hormones as examples.
Key wordsBioluminescence Binding Ligand Peptide Protein Receptor Tracer
We thank Promega Corporation for providing the plasmids encoding NanoLuc. This work was supported by the National Natural Science Foundation of China (31470767, 31270824) and the Fundamental Research Funds for the Central Universities (2000219098).
- 4.Hall MP, Unch J, Binkowski BF, Valley MP, Butler BL, Wood MG, Otto P, Zimmerman K, Vidugiris G, Machleidt T, Robers MB, Benink HA, Eggers CT, Slater MR, Meisenheimer PL, Klaubert DH, Fan F, Encell LP, Wood KV (2012) Engineered luciferase reporter from a deep sea shrimp utilizing a novel imidazopyrazinone substrate. ACS Chem Biol 7:1848–1857CrossRefPubMedPubMedCentralGoogle Scholar
- 8.McKinney M, Raddatz R (2006) Practical aspects of radioligand binding. Curr Protoc Pharmacol, Chapter 1: Unit 1.3
- 17.Zhang L, Song G, Xu T, Wu QP, Shao XX, Liu YL, Xu ZG, Guo ZY (2013) A novel ultrasensitive bioluminescent receptor-binding assay of INSL3 through chemical conjugation with Nanoluciferase. Biochimie 95:2454–2459Google Scholar