title: Current Approaches Toward Quantitative Mapping of the Interactome.
abstract: Protein-protein interactions (PPIs) play a key role in many, if not all, cellular processes. Disease is often caused by perturbation of PPIs, as recently indicated by studies of missense mutations. To understand the associations of proteins and to unravel the global picture of PPIs in the cell, different experimental detection techniques for PPIs have been established. Genetic and biochemical methods such as the yeast two-hybrid system or affinity purification-based approaches are well suited to high-throughput, proteome-wide screening and are mainly used to obtain qualitative results. However, they have been criticized for not reflecting the cellular situation or the dynamic nature of PPIs. In this review, we provide an overview of various genetic methods that go beyond qualitative detection and allow quantitative measuring of PPIs in mammalian cells, such as dual luminescence-based co-immunoprecipitation, Förster resonance energy transfer or luminescence-based mammalian interactome mapping with bait control. We discuss the strengths and weaknesses of different techniques and their potential applications in biomedical research.
authors.affiliation: Max Delbrueck Center for Molecular Medicine Berlin, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Buntru
authors.affiliation: Max Delbrueck Center for Molecular Medicine Berlin, Germany.
authors.forename: Philipp
authors.initials: P
authors.lastname: Trepte
authors.affiliation: Max Delbrueck Center for Molecular Medicine Berlin, Germany.
authors.forename: Konrad
authors.initials: K
authors.lastname: Klockmeier
authors.affiliation: Max Delbrueck Center for Molecular Medicine Berlin, Germany.
authors.forename: Sigrid
authors.initials: S
authors.lastname: Schnoegl
authors.affiliation: Max Delbrueck Center for Molecular Medicine Berlin, Germany.
authors.forename: Erich E
authors.initials: EE
authors.lastname: Wanker
keywords: BRET; BiFC; DULIP; FCCS; FRET; Interactome Mapping; LUMIER; PLA; PPI analysis; Quantification of protein-protein interactions