A split luciferase-based probe for quantitative proximal determination of Gα_q signalling in live cells.

ABSTRACT: The earlier an activation of a G protein-dependent signalling cascade at a G protein-coupled receptor (GPCR) is probed, the less amplificatory effects contribute to the measured signal. This is especially useful in case of a precise quantification of agonist efficacies, and is of paramount importance, when determining agonist bias in relation to the β-arrestin pathway. As most canonical assays with medium to high throughput rely on the quantification of second messengers, and assays affording more proximal readouts are often limited in throughput, we developed a technique with a proximal readout and sufficiently high throughput that can be used in live cells. Split luciferase complementation (SLC) was applied to assess the interaction of Gα_q with its effector phospholipase C-β3.

SUBMITTER: Littmann T 

PROVIDER: S-EPMC6249299 | biostudies-literature | 2018 Nov

REPOSITORIES: biostudies-literature