Open AccessNoninvasive imaging of protein–protein interactions in living subjects by using reporter protein complementation and reconstitution strategies
Author(s) -
Ramasamy Paulmurugan,
Yoshio Umezawa,
Sanjiv S. Gambhir
Publication year - 2002
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.242594299
Subject(s) - myod , luciferase , protein–protein interaction , bioluminescence , microbiology and biotechnology , complementation , transfection , green fluorescent protein , bimolecular fluorescence complementation , biology , myogenin , gene , genetics , myocyte , biochemistry , mutant , myogenesis
In this study we have developed bioluminescence-imaging strategies to noninvasively and quantitatively image protein-protein interactions in living mice by using a cooled charge-coupled device camera and split reporter technology. We validate both complementation and intein-mediated reconstitution of split firefly luciferase proteins driven by the interaction of two strongly interacting proteins, MyoD and Id. We use transient transfection of cells and image MyoD-Id interaction after induction of gene expression in cell culture and in cells implanted into living mice. Techniques to study protein-protein interactions in living subjects will allow the study of cellular networks, including signal transduction pathways, as well as development and optimization of pharmaceuticals for modulating protein-protein interactions.