A rapid method for detecting protein-nucleic acid interactions by protein induced fluorescence enhancement.

ABSTRACT: Many fundamental biological processes depend on intricate networks of interactions between proteins and nucleic acids and a quantitative description of these interactions is important for understanding cellular mechanisms governing DNA replication, transcription, or translation. Here we present a versatile method for rapid and quantitative assessment of protein/nucleic acid (NA) interactions. This method is based on protein induced fluorescence enhancement (PIFE), a phenomenon whereby protein binding increases the fluorescence of Cy3-like dyes. PIFE has mainly been used in single molecule studies to detect protein association with DNA or RNA. Here we applied PIFE for steady state quantification of protein/NA interactions by using microwell plate fluorescence readers (mwPIFE). We demonstrat

SUBMITTER: Valuchova S 

PROVIDER: S-EPMC5180085 | biostudies-literature | 2016 Dec

REPOSITORIES: biostudies-literature