
ForwardMAPPIT
In the basic lay-out of ForwardMAPPIT, the bait is fused to a signalling-deficient receptor chimera and the prey is tethered to a receptor fragment containing functional STAT3 recruitment sites. Bait-prey interaction reconstitutes a functional receptor complex which, upon ligand binding, initiates the JAK-STAT signalling cascade. For analysis of the interaction between designated protein pairs, the ForwardMAPPIT mode can be combined with a binary assay. For detecting new interactions, FACS or array screening assays are available. A new focus is on the development of FACS-based MAPPIT strategies to test virtually all possible point mutants of a protein against all its interactions that are detectable via MAPPIT for large-scale "edgetics" studies. Similar strategies are being developed to study very large mutant panels in multiple signaling assays in parallel.

More information:
Related publications (methods):
- Eyckerman et al. Design and application of a cytokine-receptor-based interaction trap. Nat. Cell Biol. (2001)
original paper describing the basic ForwardMAPPIT setup - Montoye et al. A systematic scan of interactions with tyrosine motifs in the erythropoietin receptor using a mammalian 2-hybrid approach. Blood (2005)
- RelayMAPPIT
describes RelayMAPPIT, a variant where a wild-type cytokine receptor is used as bait - Montoye et al. Analysis of leptin signalling in hematopoietic cells using an adapted MAPPIT strategy. FEBS Lett. (2006)
- BetaMAPPIT
describes BetaMAPPIT, a variant where STAT5 is used as signalling mediator instead of STAT3 and the prey is fused to a fragment of the beta common receptor chain instead of gp130; this modification allows MAPPIT to be used in cell types not expressing high enough endogeneous STAT3 levels, e.g. hematopoietic cells
Related publications (protocol):
Related publications (applications):
- Montoye et al. Analysis of leptin signalling in hematopoietic cells using an adapted MAPPIT strategy. FEBS Lett. (2006)
- Lavens et al. A complex interaction pattern of CIS and SOCS2 with the leptin receptor. J. Cell Sci. (2006)
- Piessevaux et al. Functional cross-modulation between SOCS proteins can stimulate cytokine signaling. J. Biol. Chem. (2006)
- Lavens et al. The C-terminus of CIS defines its interaction pattern. Biochem. J. (2007)
- Erkeland et al. Novel role of WD40 and SOCS box protein-2 in steady-state distribution of granulocyte colony-stimulating factor receptor and G-CSF-controlled proliferation and differentiation signaling. Oncogene (2007)
- Ulrichts et al. MAPPIT analysis of TLR adaptor complexes. FEBS Lett. (2007)
- Uyttendaele et al. MAPPIT analysis of STAT5, CIS and SOCS2 interactions with the growth hormone receptor. Mol. Endocrinol. (2007)
- Wauman et al. Insulin receptor substrate 4 couples the leptin receptor to multiple signaling pathways. Mol. Endocrinol. (2008)
- Ulrichts and Tavernier. MAPPIT analysis of early Toll-like receptor signalling events. Immunol. Lett. (2008)
- Piessevaux et al. Elongin B/C recruitment regulates substrate binding by CIS. J. Biol. Chem. (2008)
- Boxem et al. A protein domain-based interactome network for C. elegans early embryogenesis. Cell (2008)
- Pattyn et al. MAPPIT (MAmmalian Protein-Protein Interaction Trap) as a tool to study HIV reverse transcriptase dimerization in intact human cells. J. Virol. Methods (2008)
- Yu et al. High-quality binary protein interaction map of the yeast interactome network. Science (2008)
- Braun et al. An experimentally derived confidence score for binary protein-protein interactions. Nat. Methods (2009)
- Venkatesan et al. An empirical framework for binary interactome mapping. Nat. Methods (2009)
- Simonis et al. A high-quality high-throughput Caenorhabditis elegans protein-protein interaction map. Nat. Methods (2009)
- Lievens et al. Array MAPPIT: high-throughput interactome analysis in mammalian cells. J. Prot. Res. (2009)
- Ulrichts et al. MAPPIT (Mammalian protein-protein interaction trap) analysis of early steps in TLR signalling. Methods Mol. Biol. (2009)
- Yan et al. Abrogation of the Brd4-positive transcription elongation factor B complex by papillomavirus E2 protein contributes to viral oncogene repression. J. Virol. (2009)
- Lavens et al. Definition of the interacting interfaces of Apobec3G and HIV-1 Vif using MAPPIT mutagenesis analysis. Nucleic Acids Res. (2010)
- Ulrichts et al. Caspase-1 targets the TLR adaptor Mal at a crucial TIR-domain interaction site. J. Cell Sci. (2010)
- Simicek et al. The deubiquitylase USP33 discriminates between RALB functions in autophagy and innate immune response. Nat. Cell Biol. (2013)
- Brozzi et al. A combined "omics" approach identifies N-Myc interactor as a novel cytokine-induced regulator of IRE1 protein and c-Jun N-terminal kinase in pancreatic beta cells. J. Biol. Chem. (2014)
- Vervliet et al. Ryanodine receptors are targeted by anti-apoptotic Bcl-XL involving its BH4 domain and Lys87 from its BH3 domain. Sci. rep. (2015)
- Brozzi et al. Ubiquitin D Regulates IRE1α/c-Jun N-terminal Kinase (JNK) Protein-dependent Apoptosis in Pancreatic Beta Cells. J. Biol. Chem. (2016)
- Petta et al. The Interactome of the Glucocorticoid Receptor and Its Influence on the Actions of Glucocorticoids in Combatting Inflammatory and Infectious Diseases. Microbiol. Mol. Biol. Rev. (2016)
- Baietti et al. OTUB1 triggers lung cancer development by inhibiting RAS monoubiquitination. EMBO Mol. Med. (2016)
- Petta et al. Glucocorticoid Receptor-mediated transactivation is hampered by Striatin-3, a novel interaction partner of the receptor. Sci. Rep. (2017)
- Masschaele et al. High-Confidence Interactome for RNF41 Built on Multiple Orthogonal Assays. J. Proteome Res. (2018)
- Dumbacher et al. Modifying Rap1-signalling by targeting Pde6δ is neuroprotective in models of Alzheimer's disease. Mol. Neurodegener. (2018)