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Noninvasive imaging of protein-protein interactions in living subjects by using reporter protein complementation and reconstitution strategies.

ABSTRACT: 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.

SUBMITTER: Paulmurugan R

PROVIDER: S-EPMC137764 | biostudies-literature | 2002 Nov

REPOSITORIES: biostudies-literature

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Noninvasive imaging of protein-protein interactions in living subjects by using reporter protein complementation and reconstitution strategies.

Paulmurugan R R Umezawa Y Y Gambhir S S SS

Proceedings of the National Academy of Sciences of the United States of America 20021118 24

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 ...[more]

PMID: 12438689

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