Clostridium perfringens epsilon toxin vaccine candidate lacking toxicity to cells expressing myelin and lymphocyte protein

Variant proteins tested in this study. a The location of residues previously mutated (Y30 and Y196; ref. 20) are shown in red and the location of residues in the β-octyl-glucoside binding cleft (V72, F92, H149, V166, A168; ref. 19) mutated in this study are shown in green. Purified proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and visualised by Coomassie staining before b and after c activation with trypsin. Lane 1？=？molecular weight marker (kDa); 2？=？wild-type toxin; 3？=？Y30A-Y196A; 4？=？Y30A-Y196A-H149A; 5？=？Y30A-Y196A-A168F; 6？=？Y30A-Y196A-F92A; 7？=？Y30A-Y196A-V166A; 8？=？Y30A-Y196A-V72F; 9？=？Y30A-Y196A-H149A-A168F. Gels shown derive from the same experiment and were processed in paralle