MAPPIT modes

The MAPPIT toolbox consists of a number of related technologies which have different modes of action: forward and reverse two-hybrid or three-hybrid. Each of these MAPPIT modes aims at specific applications in the protein interaction field.


ForwardMAPPIT, the forward two-hybrid setup, allows for the detection of interactions between two proteins. This mode can be used to analyze designated protein interactions in binary assays and to identify novel interactions in FACS- or array-based screens. Read more »


In addition, a number of two-hybrid variants have been developed for specific applications. Of particular value is the possibility to incorporate a modifying enzyme in the interaction trap for the identification of modification-dependent interactions with HeteromericMAPPIT. Read more »


In the reverse mode, a signal is generated upon disruption of a protein-protein interaction. In this way, ReverseMAPPIT enables screening for proteins (mutants or fragments) or small organic molecules that interfere with a given protein interaction. Read more »


MASPIT (MAmmalian Small molecule-Protein Interaction Trap) is the three-hybrid component of the MAPPIT platform where a small organic molecule is displayed as bait, allowing for the detection of proteins that interact with these compounds. Read more »


A limitation of MAPPIT is that the interaction detection zone is restricted to the plasmamembrane environment, and that it is incompatible with full size transmembrane proteins. To overcome these restrictions KISS (for KInase Substrate Sensor) was developed, an adaptation from MAPPIT whereby the signaling incompetent receptor is replaced by the kinase domain of Tyk2, a JAK2 family member.
Since KISS uses the same prey configuration and read-out as MAPPIT, the array screening platform is transferable to the KISS assay. Read more »