Prx II and CKBB proteins interaction under physiological and thermal stress conditions in A549 and HeLa cells

Abstract

Peroxiredoxins (Prxs) are versatile enzymes that demonstrate various cell functions as peroxidases, protein chaperones, functions of signal modulators and binding partners. It is well established that Prxs can interact with multiple proteins in cells, such as ASK1, Cdk5-p35, JNK, MIF, PDGF, TKR4 and others. In this study, we attempted to evaluate a possible association between ubiquitous Prx II and ATP/ADP buffering enzyme – brain-type creatine kinase (CKBB). Our co-immunoprecipitation (Co-IP) results from the A549 and HeLa cell lysates with overexpressed HA-Prx II and Flag-CKBB have demonstrated strong association between two proteins under non-stressed conditions. This protein interaction was enhanced by the heat treatment with further HA-Prx II precipitation to the immobilized Flag-CKBB depending on the temperature increase. Temperature induced oligomerization of Prx II may contribute to the formation of Prx II conglomera­tes, which in turn, can associate with CKBB and increase signal intensities on the blotted membranes. Thus, such association and oligomerization of Prx II could take part in recovery and protection of the CKBB enzyme activity from inactivation during heat-induced stress.