Noninvasive Imaging of Protein-Protein Interactions in Living Subjects by Using Reporter Protein Complementation and Reconstitution Strategies

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 o...

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Published inProceedings of the National Academy of Sciences - PNAS Vol. 99; no. 24; pp. 15608 - 15613
Main Authors Paulmurugan, R., Umezawa, Y., Gambhir, S.S.
Format Journal Article
LanguageEnglish
Published United States National Academy of Sciences 26.11.2002
National Acad Sciences
Subjects
Animals
Biological Sciences
Bioluminescence
Cell culture techniques
Cell Line - transplantation
Cell lines
Cercopithecus aethiops
Complementation
COS Cells
Cytomegalovirus
Drug interactions
Flues
Fluorescence
Genes, Reporter
Genes, Synthetic
Genetic Complementation Test
Genetic vectors
Humans
Imaging
Inhibitor of Differentiation Protein 1
Kidney
Luciferases - analysis
Luminescent Measurements
Medical research
Mice
MyoD Protein - chemistry
MyoD Protein - metabolism
NF-kappa B - metabolism
Peptide Fragments - analysis
Peptide Fragments - genetics
Plasmids
Promoter Regions, Genetic
Protein Interaction Mapping
Proteins
Recombinant Fusion Proteins - analysis
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - physiology
Repressor Proteins
Reproducibility of Results
Rodents
Transcription Factors - chemistry
Transcription Factors - metabolism
Transfection
Tumor Cells, Cultured
Tumor Necrosis Factor-alpha - pharmacology
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Summary: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.
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Communicated by Michael E. Phelps, University of California, Los Angeles, CA
To whom correspondence should be addressed at: Crump Institute for Molecular Imaging, University of California School of Medicine, B3–399A BRI, 700 Westwood Plaza, Los Angeles, CA 90095-1770. E-mail: sgambhir@mednet.ucla.edu.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.242594299