Chapter title |
MAPPIT, a Mammalian Two-Hybrid Method for In-Cell Detection of Protein-Protein Interactions
|
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Chapter number | 29 |
Book title |
Protein-Protein Interactions
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Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-2425-7_29 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2424-0, 978-1-4939-2425-7
|
Authors |
Irma Lemmens, Sam Lievens, Jan Tavernier, Lemmens, Irma, Lievens, Sam, Tavernier, Jan |
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. |
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