ArrayMAPPIT: a screening platform for human protein interactome analysis.
Mammalian protein-protein interaction trap (MAPPIT) is a two-hybrid technology to identify and characterize interactions of proteins with other proteins or organic molecules in living mammalian (human) cells. The method relies on complementation of a modified cytokine receptor complex. Protein interaction restores the signalling competence of the complex, which is monitored through the activation of a reporter gene. Here, we describe a protocol that has been recently developed to increase the utility of MAPPIT as a tool to identify novel interactions. In the ArrayMAPPIT assay, a collection of prey proteins which is arrayed in high-density microtiter plates is efficiently screened for interaction partners using reverse transfection into a bait-expressing cell pool.
Department of Medical Protein Research, VIB and Department of Biochemistry, Ghent University, Ghent, Belgium.
Sam
S
Lievens
Nele
N
Vanderroost
Dieter
D
Defever
José
J
Van der Heyden
Jan
J
Tavernier
D000818:Animals; D006801:Humans; D010957:Plasmids / Q000235:genetics; D025941:Protein Interaction Mapping / Q000295:instrumentation* / Q000379:methods; D011506:Proteins / Q000032:analysis* / Q000378:metabolism*; D013997:Time Factors; D014162:Transfection
D011506:Proteins