2D-ELECTROPHORESIS IN PROTEIN SPECTRUM CONSTRUCTION FOR EXOPROTEIN FRACTIONS OF PLAGUE AND CHOLERA AGENTS

Abstract

Objective of the study is to apply 2D-electrophoresis for imaging protein spectrum of exoprotein fractions, as well as identification and comparison of biomarker expression of the key pathogenicity factors in plague and cholera agents. Materials and methods. 2D gels of exoprotein fractions were obtained on the model of Vibrio cholerae Inaba 569B, V. cholerae El Tor M888Ctx+ and Ctx– strains, and also Y. pestis EV NIIEG strain. Capsular antigen F1 and cholera toxin were used as biomarkers of major exoproteins of plague and cholera agents. Results and conclusions. While studying cholera toxin through 2D-electrophoresis on IPG strips with pH 3–10 gradient, the antigen fell into a number of protein spots, two of which were close to A1 domain parameters (MW 20.309 kDa and pI 6.51) and B-subunit monomer (MW 11.091 kDa and pI 7.68). Comparative protein spectrum analysis of the exoprotein fractions of V. cholerae Inaba 569B, V. cholerae El Tor M888 Ctx+ and Ctx– has revealed the spots with similar parameters, excluding the strain V. cholerae M888 Ctx– . Investigation of 2D gels of capsular antigen and exoprotein fractions of Y. pestis EV strain has demonstrated that protein spots corresponding to the parameters of the subunit form and diameter of the antigen are present in 37 °C culture patterns. 28 °C culture sample contains F1 subunit in a far lesser concentrations. On the model of toxigenic V. cholerae Inaba 569B strain, isogenic system of V. cholerae El Tor M888 Ctx+ and Ctx– strains, and Y. pestis EV NIIEG strain, high efficiency of 2D-electrophoresis in protein spectrum construction for exoprotein fractions of the cultures, as well as identification and comparison of biomarker expression of major exoproteins of plague and cholera agents is established.

Keywords

• 2d-electrophoresis
• exoproteins
• vibrio cholerae
• yersinia pestis