Visualizing protein-protein interactions in living animals

Gary D. Luker, Vijay Sharma, David Piwnica-Worms

Research output: Contribution to journalArticle

44 Scopus citations

Abstract

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.

Original languageEnglish
Pages (from-to)110-122
Number of pages13
JournalMethods
Volume29
Issue number1
DOIs
StatePublished - Jan 1 2003

Keywords

  • Green fluorescent protein
  • Herpes simplex virus
  • Large T antigen
  • Molecular imaging
  • Positron emission tomography
  • Protein-protein interactions
  • Reporter genes
  • Thymidine kinase
  • Two-hybrid system
  • p53

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