Eue I, Kauschke E, Mohrig W, Cooper E L
Universität Münster, Institut für Experimentelle Dermatologie, Germany.
Dev Comp Immunol. 1998 Jan-Feb;22(1):13-25. doi: 10.1016/s0145-305x(97)00049-9.
Hemolytic activity in coelomic fluid of Eisenia fetida (ECF) is due to three proteins H1, H2, H3 with molecular weights of 46, 43 and 40 kD, respectively. These proteins were isolated by preparative PAGE. H1 and H2 were shown to be stable in SDS and alpha-2-ME whereas H3 splits into two fragments with molecular weights of 18 and 21 kD after SDS treatment. IEF indicates that each protein consists of different isoforms with pIs between 5.1 and 6.2 H3 was demonstrated to be a bifunctional protein that can lyse and agglutinate erythrocytes. At 56 degrees C hemolytic activity of all three proteins was inactivated, but the agglutination activity of H3 was stable. Intracoelomic injection of erythrocytes reduced the number of hemolysins from three to two. Monospecific antisera were raised against the isolated hemolysins H1,2 and 3. The use of these antibodies and of carbohydrates as inhibitors of the biological activity of the molecules demonstrates the close structural relationship of agglutinins and hemolysins in the CF of E. fetida.
赤子爱胜蚓体腔液（ECF）中的溶血活性归因于三种蛋白质H1、H2、H3，其分子量分别为46、43和40kD。这些蛋白质通过制备型聚丙烯酰胺凝胶电泳进行分离。H1和H2在十二烷基硫酸钠（SDS）和α-2-巯基乙醇（α-2-ME）中表现稳定，而H3在SDS处理后分解为分子量分别为18和21kD的两个片段。等电聚焦（IEF）表明每种蛋白质由不同的同工型组成，其等电点在5.1至6.2之间。H3被证明是一种双功能蛋白，能够裂解和凝集红细胞。在56℃时，所有三种蛋白质的溶血活性均失活，但H3的凝集活性保持稳定。向体腔内注射红细胞可使溶血素的数量从三种减少到两种。针对分离出的溶血素H1、H2和H3制备了单特异性抗血清。使用这些抗体以及糖类作为分子生物活性的抑制剂，证明了赤子爱胜蚓体腔液中凝集素和溶血素之间存在密切的结构关系。
