MrkD adhesin of Klebsiella pneumoniae expression, purification and analysis of adhesive activity

Abstract:

Abstract: [Objective] The mrkD gene encodes the adhesin which mediates Klebsiella pneumoniae to adhere human respiratory tissue.We aimed to analyze the adhesion mechanism and adhesion block funcntion of MrkD adhesin. [Methods] The recombinant glutathione-S-transferase(GST)-tagged adhesive protein(MrkD) was expressed in E. coli and was purified to homogeneity using GST affinity chromatography. The GST tag was cut by thrombin to obtain the MrkD protein that was identified by SDS-PAGE and Western blot. The adhesive activity of MrkD was examined by adhesive experiments and the binding site was observed by laser confocal microscopy. [Results] The adherent activity of Klebsiella pneumoniae was significantly inhibited by the MrkD. These experimental data demonstrated that the MrkD inhibited the adhesion of Klebsiella pneumoniae. [Conclusion] Our results suggest that MrkD adhesin contains the adhesion epitopes.The future work will be carried out to identify the epitopes and characterize them, then to optimize the combination presentation of these epitopes to develop an efficient vaccine for Klebsiella pneumoniae.