Reverse MAPPIT detects disruptors of protein-protein interactions in human cells.
Reverse mammalian protein-protein interaction trap (MAPPIT) is a mammalian reverse two-hybrid technology. The method is adapted from the forward MAPPIT technique, a two-hybrid complementation system in which the interaction between a bait-fusion protein and a prey-fusion protein restores ligand-dependent cytokine receptor signaling. In the reverse mode described in detail here, a positive readout is generated on disruption of the designated protein-protein interactions. Reverse MAPPIT functions in intact human cells, facilitating simultaneous analysis of disruption, toxicity and permeability of the tested compounds, making it particularly suitable for screening for molecules that target therapeutically interesting protein-protein interactions or for mapping the interaction interface between proteins. The total handling time of a typical reverse MAPPIT experiment is approximately 9 h and is spread over 4-5 d.
Lemmens
Irma
I
Department of Medical Protein Research, Faculty of Medicine and Health Sciences, Flanders Interuniversity Institute for Biotechnology, VIB09, Ghent University, A. Baertsoenkaai 3, B-9000 Ghent, Belgium.
Lievens
Sam
S
Eyckerman
Sven
S
Tavernier
Jan
J
D002478:Cells, Cultured; D006801:Humans; D008024:Ligands; D018121:Receptors, Cytokine / Q000378:metabolism; D020798:Two-Hybrid System Techniques
D008024:Ligands; D018121:Receptors, Cytokine