The core technology of Sungkyunkwan University is to provide a method for in vitro phosphorylation of TRAP. The method comprises a step in vitro mixing a phosphorylation buffer, oxidizing ions of a transition metal such as iron (Fe2+), copper (Cu2+) and zinc (Zn2+), and ATP with TRAP purified from cells.
Background and unmet needs: Rap and the TRAP proteins of Staphylococcus are very important in initiating the host infection via the signal transduction. Especially, the TRAP phosphorylation is essential in determining the staphylococcal infection. Therefore, it is expected that inhibitors against the TRAP phosphorylation can be developed into a novel antibiotic to prevent the infection of Staphylococcus and other bacteria having similar proteins to the TRAP.
Discovery and Achievements: The inventors have elucidated that the TRAP is a kinase phosphorylating itself even in the absence of a RAP protein and utilizes specifically oxidative metal ions such as iron, copper and zinc. Based upon this finding, we have developed a novel method for phosphorylating purified TRAPs in vitro. We have also attempted to provide a method for screening inhibitors against the TRAP phosphorylation based upon this technique, and completed this technique successfully.