Journal
PROTEOMICS
Volume 8, Issue 6, Pages 1179-1196Publisher
WILEY-BLACKWELL
DOI: 10.1002/pmic.200700802
Keywords
bioluminescence; fluorescence; imaging; protein-protein interaction; resonance energy transfer
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This review aims to provide an overview of current optical procedures used in functional proteomics, investigating protein localization, protein-protein interaction, intracellular signaling events, and second messenger generation in living cells. Reporter assays using proteins tagged with fluorescent or bioluminescent moieties are discussed. Recently, intracellular biosensor assays, flow cytometry-based techniques (fluorescent cell barcoding), as well as transfected cell microarray assays involving RNA interference coupled with automated imaging were introduced and have been adopted as screening platforms for annotating small molecules, investigating signaling events, or in phenotype analysis. These novel methodological advances include improved image acquisition and processing techniques and help linking in vitro observations to in vivo processes. In addition, the acquired data are increasingly quantitative In nature and will therefore pave the way for modeling of signaling cascades and other complex cellular events, an important step toward systems biology.
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