MAPPIT (mammalian protein-protein interaction trap) analysis of early steps in toll-like receptor signalling.

The mammalian protein-protein interaction trap (MAPPIT) is a two-hybrid technique founded on type I cytokine signal transduction. Thereby, bait and prey proteins are linked to signalling deficient cytokine receptor chimeras. Interaction of bait and prey and ligand stimulation restores functional JAK (Janus kinase)-STAT (signal transducers and activators of transcription) signalling, ...
which ultimately leads to the transcription of a reporter or marker gene under the control of the STAT3-responsive rPAP1 promoter. In the subsequent protocol, we describe the use of MAPPIT to study early events in Toll-like receptor (TLR) signalling. We here demonstrate a "signalling interaction cascade" from TLR4 to IRAK-1.
Mesh Terms:
Animals, Biological Assay, Cell Line, Genetic Vectors, Humans, Protein Binding, Signal Transduction, Time Factors, Toll-Like Receptors, Transgenes
Methods Mol. Biol.
Date: Apr. 21, 2009
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