SpringerProtocols: Abstract: MAPPIT (Mammalian Protein?Protein Interaction Trap) Analysis of Early Steps in Toll-Like Receptor Signalling

9. MAPPIT (Mammalian Protein–Protein Interaction Trap) Analysis of Early Steps in Toll-Like Receptor Signalling

By: Peter Ulrichts³, Irma Lemmens³, Delphine Lavens⁴, Rudi Beyaert³, Jan Tavernier³

Abstract

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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.

Affiliation(s): (3) Department of Medical Protein Research, Ghent University – VIB, Ghent, Belgium
(4) Department of Biochemistry, Faculty of Medicine and Health Sciences, Ghent University, A. Baertsoenkaai 3, B-9000 Ghent, Belgium

Book Title: Toll-Like Receptors: Methods and Protocols

Series: Methods in Molecular Biology | Volume: 517 | Pub. Date: Jun-01-2009 | Page Range: 1-12 | DOI: 10.1007/978-1-59745-541-1_9

Subject: Immunology

Key Words: MAPPIT - Two-hybrid - JAK-STAT - Cytokine signal transduction - TLR

References

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