Visualizing protein-protein interactions in living animals.
A variety of techniques have been developed to analyze protein-protein interactions in vitro and in cultured cells. However, these methods do not determine how protein interactions affect and are regulated by physiologic and pathophysiologic conditions in living animals. This article describes methodology for detecting and quantifying protein interactions in living mice, using an inducible two-hybrid system developed for positron emission tomography (PET) imaging. We discuss the methods to establish stably transfected cells with components of the imaging system, create tumor xenografts, synthesize PET radiopharmaceuticals used to visualize the imaging reporter, perform microPET imaging, and analyze data from imaging studies. Development and application of technologies for molecular imaging of protein-protein interactions in vivo should enable researchers to investigate intrinsic binding specificities of proteins during normal development and disease progression as well as aid drug development through direct interrogation of molecular targets within intact animals.
Luker
Gary D
GD
Department of Molecular Biology, Molecular Imaging Center, Mallinckrodt Institute of Radiology, Washington University Medical School, 510 South Kingshighway Boulevard, 63110, St. Louis, MO, USA.
Sharma
Vijay
V
Piwnica-Worms
David
D
D000818:Animals; D002478:Cells, Cultured; D017930:Genes, Reporter; D008856:Microscopy, Fluorescence / Q000379:methods*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D014055:Tomography, Emission-Computed / Q000379:methods*; D014162:Transfection; D020798:Two-Hybrid System Techniques
D011994:Recombinant Proteins