Quantitation of protein-protein interactions by thermal stability shift analysis.

ABSTRACT: Thermal stability shift analysis is a powerful method for examining binding interactions in proteins. We demonstrate that under certain circumstances, protein-protein interactions can be quantitated by monitoring shifts in thermal stability using thermodynamic models and data analysis methods presented in this work. This method relies on the determination of protein stabilities from thermal unfolding experiments using fluorescent dyes such as SYPRO Orange that report on protein denaturation. Data collection is rapid and straightforward using readily available real-time polymerase chain reaction instrumentation. We present an approach for the analysis of the unfolding transitions corresponding to each partner to extract the affinity of the interaction between the proteins. This method does

SUBMITTER: Layton CJ 

PROVIDER: S-EPMC3189529 | biostudies-literature | 2011 Aug

REPOSITORIES: biostudies-literature