In ReverseMAPPIT, interference with a protein-protein interaction results in dissociation of an inhibitor from an active cytokine receptor complex. Therefore, the bait is fused to a receptor chimera in which the tyrosine involved in STAT3 docking is retained, making it fully functional in STAT signalling. The prey is fused to a phosphatase domain that inhibits the signalling pathway. Interaction between this inhibitory prey and the bait therefore blocks signalling and concomitant reporter induction. Disruption of the interaction, either by a competing protein or a small organic molecule, restores signalling and thus leads to a positive readout. This readout is one of the binary assays based on either STAT3 dependent reporter induction or direct measurement of STAT3 activation.

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