Protein complementation assays: Approaches for the in vivo analysis of protein interactions

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.

Abbreviations: 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

Keywords: Protein-fragment complementation assay, Protein interaction, Protein inhibition, Bimolecular fluorescence complementation