Analysis of Protein–DNA Interactions Using Surface Plasmon Resonance

Abstract

Protein–DNA interactions are required for access and protection of the genetic information within the cell. Historically these interactions have been studied using genetic, biochemical, and structural methods resulting in qualitative or semiquantitative interaction data. In the future the focus will be on high quality quantitative data to model a huge number of interactions forming a specific network in system biology approaches.

Toward this aim, BIAcore introduced in 1990 the first commercial machine that uses surface plasmon resonance (SPR) to study the real-time kinetics of biomolecular interactions. Since then systems have been developed to allow for robust analysis of a multitude of protein–DNA interactions. Here we provide a detailed guide for protein–DNA interaction analysis using the BIAcore, starting with a description of the SPR technology, giving recommendations on preliminary studies, and finishing with extensive information on quantitative and qualitative data analysis. One focus is on cooperative protein–DNA interactions, where proteins interact with each other to modulate their binding specificity or affinity. The BIAcore has been used for the last 14 years to study protein–DNA interactions; our literature review focuses on some high quality studies describing a wide range of experimental uses, covering simple 1:1 interactions, analysis of complicated multiprotein–DNA interaction systems, and analytical uses.