Fig. 7
From: NanoBRET in C. elegans illuminates functional receptor interactions in real time
Scheme of potential receptor internalisation tracked with enhanced bystander BRET. 1 The energy donor (Nluc C-terminally fused to NPR-11) generates a bystander BRET with an energy acceptor (mNeonGreen fused to the CAAX box of let-60) located at the inner membrane upon furimazine application. In the presence of the agonist (FLP-34-1), the receptor undergoes endocytosis (2), lowering the number of available donor molecules at the membrane that are able to transfer energy to the acceptor. This effect shifts the ratio of luminescence and fluorescence and subsequently, yields a lower BRET window. The receptor is then either degraded in lysosomes (3a) or recycled in recycling-endosomes and transported back to the membrane. The latter one increases the bystander BRET again due to the close proximity of donor and acceptor (3b). Image created with BioRender.com