Binding of Clostridium perfringens 125I-labeled delta-toxin to erythrocytes.
Hemolytically active, 125I-labeled delta-toxin from Clostridium perfringens was used to study the binding of this cytolysin to sheep, goat, human, rabbit, horse, mouse, and guinea pig erythrocytes. The extent of toxin binding was correlated with the known hemolytic specificity of the toxin. Detailed studies of the binding were carried out on sheep erythrocytes which showed the highest sensitivity to lysis by delta-toxin. Simultaneous determination of toxin binding and release of intracellular 86Rb+ and hemoglobin suggested that toxin binding and membrane damage were separate sequential events. Toxin binding was rapid (2-5 min) and temperature-dependent. The extent of binding was temperature-independent. Binding was saturable, specific, relatively tight (Ka = 4.4 X 10(8) M-1) and largely irreversible. A single type of binding site (7,000/sheep erythrocyte) was found. Cell-bound toxin was extractable by chaotropic ions. Preincubation of the toxin with N-acetylgalactosaminyl-[N-acetylneuraminyl]-galactosylglucosylceramide (GM2 ganglioside) inhibited both binding and hemolysis. Toxin binding was affected by pretreatment of sheep erythrocytes with pronase but not with trypsin or chymotrypsin. Cell treatment with neuraminidase prevented toxin binding by 30%. Preincubation of the toxin with specific immune sera blocked its binding on target cells. It is suggested that GM2 ganglioside, a more complex membrane component containing this glycolipid or a structurally related molecule is the binding site for delta-toxin on the surface of sensitive erythrocytes.
D000818:Animals; D001427:Bacterial Toxins / Q000378:metabolism*; D004912:Erythrocytes / Q000378:metabolism*; D006041:Goats; D006168:Guinea Pigs; D006460:Hemolysin Proteins / Q000378:metabolism*; D006736:Horses; D006801:Humans; D007457:Iodine Radioisotopes; D007700:Kinetics; D051379:Mice; D011817:Rabbits; D012756:Sheep; D013045:Species Specificity; D013696:Temperature
D001427:Bacterial Toxins; C029352:Clostridium perfringens delta-toxin; D006460:Hemolysin Proteins; D007457:Iodine Radioisotopes