Studies on the biological properties of coelomic fluid of sea urchin 1. Naturally occurring hemolysin in sea urchin

Abstract

1. 1. Biological properties, especially hemolytic activity, of coelomic fluid of sea urchin were investigated. 2. 2. Dialyzed coelomic fluid preparation obtained from Anthocidaris crassispina was found to have hemolytic activity against rabbit, mouse and human erythrocytes, but not against erythrocytes of other species of animals so far examined, i.e. rat, guinea pig, goat, sheep, dog, pig, hen, snake and frog. A similar hemolysis spectrum was obtained with a coelomic fluid preparation from Pseudocentrotus depressus, while coelomic fluid preparation from Hemicentrotus pulcherrimus showed a capacity to lyse erythrocytes of rat and guinea pig, besides those of rabbit, mouse and man. 3. 3. The hemolytic activity of coelomic fluid preparation was greatly enhanced by the addition of Ca 2+ to the medium, but not by Mg 2+. Addition of EDTA, even in the presence of Ca 2+, resulted in a loss of potency. 4. 4. The degree of hemolysis induced by coelomic fluid preparation was dependent on concentration and temperature of incubation. The kinetic analysis as a function of concentration indicated that the hemolysis curve resembles those of saponin and streptolysin O. 5. 5. The hemolytic factor in coelomic fluid preparation was so labile that complete inactivation was effected after 30 min of incubation at 56°C, and after storage for 24 h at pH 11.3. 6. 6. The active factor may be not a saponin but a protein or a protein-like substance, since its activity was abolished irreversibly by treatment with trypsin or 2-mercaptoethanol.