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How The PathHunter® β-Arrestin GPCR Assays Work
In this system, the GPCR is fused in frame with the small, 42 amino acid fragment
of β-gal called ProLink™ and co-expressed in cells stably expressing a fusion protein
of β-Arrestin and the larger, N-terminal deletion mutant of β-gal (called enzyme
acceptor or EA). Activation of the GPCR stimulates binding of β-Arrestin to the
ProLink-tagged GPCR and forces complementation of the two enzyme fragments,
resulting in the formation of an active β-gal enzyme. This action leads to an increase
in enzyme activity that can be measured using chemiluminescent PathHunter®
Detection Reagents.
PathHunter® β-Arrestin Assay Principle.
Activation of the ProLink-tagged GPCR results in β-Arrestin recruitment and formation of a functional
enzyme capable of hydrolyzing substrate and generating a chemiluminescent signal.
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