Identification of an interaction between EI and a histidine kinase-response regulator hybrid protein in Gluconobacter oxydans

Abstract

Gluconobacter oxydans may contain an incomplete phosphoenolpyruvate: carbohydrate phosphotransferase system consisting of three components--EI, HPr and EIIA, while the function of individual members of the system remains unknown. In this research, a specific interaction between EI and a histidine kinase-response regulator hybrid protein was screened by yeast two-hybrid assay, and the interaction was further identified with GST pull-down assay and bimolecular fluorescence complementation assay in vitro and in vivo, respectively. As the histidine kinase-response regulator hybrid protein serves as a member of two-component system in G. oxydans, its interaction with EI implied that PTS may play certain roles in bacteria under stress.

Keywords: Bimolecular fluorescence complementation; GST pull-down; Gluconobacter oxydans; Phosphoenolpyruvate: carbohydrate phosphotransferase system; Two-component system; Yeast two-hybrid assay.