Identification and characterization of the Aeromonas sobria hemolysin glycoprotein receptor on Intestine 407 cells

Aeromonas sobria hemolysin is important in the pathogenesis of diarrhoea ca used by this enteropathogenic bacterium. By immunoprecipitation analysis us ing hemolysin and anti-hemolysin antibody, a 66 kDa protein (p66) was ident ified as a receptor for A. sobria hemolysin on Intestine 407 cells. Treatme nt of p66 with N-glycosidase F reduced the apparent sized of p66 to 60 kDa on SDS-polyacrylamide gels, p66, released from Intestine 407 cells followin g incubation with phosphatidylinositol-specific phospholipase C (PI-PLC) tr eatment, bound A. sobria hemolysin. Thus treatment of Intestine 407 cells w ith PI-PLC resulted in the remarkable decrease of the sensitivity to A. sob ria hemolysin. These results are consistent with the hypothesis that p66, t he binding protein for A. sobria hemolysin, is a glycosylphosphatidylinosit ol-anchored glycoprotein expressed on the surface of Intestine 407 cells an d probably plays a role as a receptor for A. sobria hemolysin on the intest inal cells.