Abstract
Extracellular products of Corynebacterium hemolyticum, C. ovis and C. ulcerans were purified by methanol precipitation and ion exchange chromatography in CM-Sephadex. Three different activities were determined i.e. dermonecrotoxin, phospholipase D (phosphatidylcholine phosphatidohydrolase EC 3.1.4.4) and inhibition of staphylococcal β lysin. All these activities were found in the same fraction. Sheep and rabbit erythrocytes were treated with partially purified toxin and the hemolytic activities of staphylococcal α and β lysins and phospholipase C (phosphatidylcholine cholinephosphohydrolase EC 3.1.4.3) of Clostridium perfringens were estimated quantitatively. Sheep erythrocytes were completely protected against the hemolytic action of staphylococcal α and β lysins. Excess of the other hemolytic agents was needed to cause hemolysis in treated erythrocytes, namely 4 HU of α lysin in rabbit erythrocytes, 256 HU of phospholipase C in sheep erythrocytes and 16 HU of amboceptor in immune hemolysis. Sphingomyelins form the major part of phospholipids in the cell membrane of sheep erythrocytes and are hydrolyzed by the corynebacterial phospholipase D which is probably the reason for the protective action of the toxin.
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