title: LuMPIS: luciferase-based MBP-pull-down protein interaction screening system.
abstract: Analyzing the putative interaction partners of an individual protein is one approach to elucidate its function. In the LuMPIS protocol, bait and prey proteins are expressed with N-terminal maltose binding protein (MBP)- and eGFP-luciferase (eGFP-luc) tags, respectively. Positive protein-protein interactions (PPIs) can be detected after pull-down of the MBP-tagged prey protein using amylose beads followed by the bioluminescence detection of the bound eGFP-luc-tagged bait protein. The LuMPIS technology offers the following advantages: the PPIs are detected in the mammalian cell context, the use of two long protein tags (i.e., MBP and eGFP-luc) increases the expression levels of genes whose gene expression levels are known to be frequently impaired, the use of amylose beads for pull-down is much more economic as compared to sepharose beads in combination with monoclonal antibodies and finally, the use of an eGFP-luc-tag enables the qualitative control of transfection efficiencies by fluorescence microscopy prior to starting the assay.
authors: Department of Virology, Max von Pettenkofer-Institute, Pettenkoferstr. 9a, Münich 80336, Germany. vizoso@mvp.uni-muenchen.de
authors: María G Vizoso
authors: MG
authors: Pinto
authors: Armin
authors: A
authors: Baiker
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000302:isolation & purification; D006801:Humans; D049410:Luciferases, Renilla / Q000737:chemistry* / Q000302:isolation & purification; D058619:Maltose-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification; D014162:Transfection; D014764:Viral Proteins / Q000737:chemistry
chemical_list: D058619:Maltose-Binding Proteins; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D049410:Luciferases, Renilla