Methods for the detection and analysis of protein-protein interactions

Research output: Contribution to journalReview article

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.

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Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Industrial Biotechnology
  • Immunology in the medical area

Keywords

  • affinity tags, protein-protein interactions, purification, protein, protein complexes
Original languageEnglish
Pages (from-to)2833-2842
JournalProteomics
Volume7
Issue number16
Publication statusPublished - 2007
Publication categoryResearch
Peer-reviewedYes