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 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.
Paulmurugan
R
R
Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, University of California Los Angeles-Jonsson Comprehensive Cancer Center, 90095-1770, USA.
Umezawa
Y
Y
Gambhir
S S
SS
D000818:Animals; D019556:COS Cells; D002460:Cell Line / Q000637:transplantation; D002522:Chlorocebus aethiops; D003587:Cytomegalovirus; D017930:Genes, Reporter; D005813:Genes, Synthetic; D005816:Genetic Complementation Test; D006801:Humans; D051798:Inhibitor of Differentiation Protein 1; D007668:Kidney; D008156:Luciferases / Q000032:analysis*; D008163:Luminescent Measurements; D051379:Mice; D017570:MyoD Protein / Q000737:chemistry / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism; D010446:Peptide Fragments / Q000032:analysis / Q000235:genetics; D011401:Promoter Regions, Genetic; D025941:Protein Interaction Mapping; D011993:Recombinant Fusion Proteins / Q000032:analysis* / Q000235:genetics / Q000502:physiology; D012097:Repressor Proteins; D015203:Reproducibility of Results; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism*; D014162:Transfection; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
C497453:ID1 protein, human; C497455:Idb1 protein, mouse; D051798:Inhibitor of Differentiation Protein 1; D017570:MyoD Protein; D016328:NF-kappa B; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D014157:Transcription Factors; D014409:Tumor Necrosis Factor-alpha; D008156:Luciferases