title: MAPPIT, a mammalian two-hybrid method for in-cell detection of protein-protein interactions.
abstract: MAPPIT (MAmmalian Protein-Protein Interaction Trap) is a two-hybrid technology that facilitates the detection and analysis of interactions between proteins in living mammalian cells. The system is based on type 1 cytokine receptor signaling. The bait protein of interest is fused to a chimeric signaling-deficient cytokine receptor, the signaling competence of which is restored upon recruitment of a prey protein that is coupled to a functional cytokine receptor domain. MAPPIT exhibits an excellent signal-to-noise ratio, detects a wide variety of protein-protein interactions (PPIs) including transient and indirect interactions, and has been shown to be highly complementary to other two-hybrid methods with respect to the interactions it can detect. Variants of the method were developed to allow large-scale PPI screening, mapping of protein interaction interfaces, PPI inhibitor screening and drug profiling. This chapter describes a basic 4-day MAPPIT protocol for the analysis of interaction between two designated proteins.
authors: Department of Medical Protein Research, VIB, A. Baertsoenkaai 3, 9000, Ghent, Belgium.
authors: Irma
authors: I
authors: Lemmens
authors: Sam
authors: S
authors: Lievens
authors: Jan
authors: J
authors: Tavernier
mesh_terms: D000818:Animals; D002460:Cell Line; D016207:Cytokines / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008322:Mammals; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D018121:Receptors, Cytokine / Q000737:chemistry / Q000378:metabolism*; D015398:Signal Transduction / Q000235:genetics; D059629:Signal-To-Noise Ratio; D020798:Two-Hybrid System Techniques
chemical_list: D016207:Cytokines; D018121:Receptors, Cytokine