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Protein complementation assays: Approaches for the in vivo analysis of protein interactions
Authors:Montse Morell  Francesc X Avilés
Institution:Institut de Biotecnologia i de Biomedicina and Departament de Bioquímica i Biologia Molecular, Facultat de Biociències, Universitat Autònoma de Barcelona, E-08193 Bellaterra, Spain
Abstract:The in vivo identification and characterization of protein-protein interactions (PPIs) are essential to understand cellular events in living organisms. In this review, we focus on protein complementation assays (PCAs) that have been developed to detect in vivo protein interactions as well as their modulation or spatial and temporal changes. The uses of PCAs are increasing, spanning different areas such as the study of biochemical networks, screening for protein inhibitors and determination of drug effects. Emphasis is given to approaches that rely on signals of spectroscopic nature (i.e. fluorescence or luminescence), the ones that are more directly related to bioimaging.
Keywords:BIFC  bimolecular fluorescence complementation  PCA  protein-fragment complementation assay  GFP  green fluorescent protein  FP  fluorescent protein  DHFR  dihydrofolate reductase  PPI  protein-protein interaction  RET  resonance energy transfer  BIFC-RET  BIFC-based resonance energy transfer  FC  flow cytometry  AP/MS  affinity purification followed by mass spectrometry  Y2H  yeast two-hybrid
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