Methods for the detection and analysis of protein-protein interactions

Summary, in English

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.

• Author: Tord Berggård; Sara Linse; Peter James
• Department/s: Department of Chemistry; Departments at LTH; Department of Immunotechnology
• Publishing year: 2007
• Language: English
• Pages: 2833-2842
• Publication/Series: Proteomics
• Volume: 7
• Issue: 16
• Links: http://dx.doi.org/10.1002/pmic.200700131
• Document type: Journal article review
• Publisher: John Wiley & Sons Inc.
• Topic: Industrial Biotechnology; Immunology in the medical area
• Keywords: affinity tags; protein-protein interactions; purification; protein; protein complexes
• Status: Published
• ISBN/ISSN/Other: ISSN: 1615-9861