title: Methods for the detection and analysis of protein-protein interactions.
abstract: A large number of methods have been developed over the years to study protein-protein interactions. Many of these techniques are now available to the nonspecialist researcher thanks to new affordable instruments and/or resource centres. A typical protein-protein interaction study usually starts with an initial screen for novel binding partners. We start this review by describing three techniques that can be used for this purpose: (i) affinity-tagged proteins (ii) the two-hybrid system and (iii) some quantitative proteomic techniques that can be used in combination with, e.g., affinity chromatography and coimmunoprecipitation for screening of protein-protein interactions. We then describe some public protein-protein interaction databases that can be searched to identify previously reported interactions for a given bait protein. Four strategies for validation of protein-protein interactions are presented: confocal microscopy for intracellular colocalization of proteins, coimmunoprecipitation, surface plasmon resonance (SPR) and spectroscopic studies. Throughout the review we focus particularly on the advantages and limitations of each method.
authors.affiliation: Department of Biophysical Chemistry, Lund University, Lund, Sweden. tord.berggard@bpc.lu.se
authors.forename: Tord
authors.initials: T
authors.lastname: Berggård
authors.forename: Sara
authors.initials: S
authors.lastname: Linse
authors.forename: Peter
authors.initials: P
authors.lastname: James
mesh_terms: D002846:Chromatography, Affinity; D030562:Databases, Protein; D047468:Immunoprecipitation; D018613:Microscopy, Confocal; D011485:Protein Binding; D011506:Proteins / Q000302:isolation & purification / Q000378:metabolism*; D040901:Proteomics; D020349:Surface Plasmon Resonance; D020798:Two-Hybrid System Techniques
chemical_list: D011506:Proteins