Purification and partial characterization of a non-O1 Vibrio cholerae hemolysin that cross-reacts with thermostable direct hemolysin of Vibrio parahaemolyticus

Abstract

A newly identified hemolysin (NAG-rTDH), which is related to the thermostable direct hemolysin (Vp-TDH) of Vibrio parahaemolyticus produced by non-O1 Vibrio cholerae, was studied. NAG-rTDH was purified by successive column chromatographies on DEAE-cellulose and an immunoaffinity column coupled with anti-Vp-TDH immunoglobulin. The molecular weight of NAG-rTDH was estimated as 18,500, similar to that of Vp-TDH, as judged by sodium dodecyl sulfate slab gel electrophoresis, but its charge or molecular shape was different, judging from its electrophoretic mobility. The lytic activities of NAG-rTDH on erythrocytes of most animals were essentially similar to those of Vp-TDH, but that on sheep erythrocytes was different. The hemolytic activity of NAG-rTDH was stable on heating at 100 degrees C for 10 min, as was that of Vp-TDH. Immunological cross-reactivity between NAG-rTDH and Vp-TDH was demonstrated by both the Ouchterlony test and the neutralization test. Thus, we conclude that non-O1 V. cholerae produce a new type of hemolysin that is similar but not identical to the thermostable direct hemolysin of V. parahaemolyticus.