Isolation and characterization of earthworm hemolysins and agglutinins

• Published in: Developmental and comparative immunology Vol. 22; no. 1; pp. 13 - 25
• Main Authors: Eue, Ines, Kauschke, Ellen, Mohrig, Werner, Cooper, Edwin L.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Ltd 1998
• Subjects: Agglutinins - immunology; Agglutinins - isolation & purification; Animals; Body Fluids - immunology; Carbohydrates - pharmacology; Coelomic fluid; Cross Reactions; Earthworms; Eisenia fetida; Glycoproteins - pharmacology; Hemagglutination - drug effects; Hemagglutinin; Hemolysin; Hemolysin Proteins - immunology; Hemolysin Proteins - isolation & purification; Hemolysis - drug effects; IEF; Oligochaeta - immunology; PAGE; Western blotting; Hemagglutinin; PAGE; Eisenia fetida; IEF; Earthworms; Hemolysin; Coelomic fluid; Western blotting
• ISSN: 0145-305X; 1879-0089
• DOI: 10.1016/S0145-305X(97)00049-9

Hemolytic activity in coelomic fluid of
Eisenia fetida (ECF) is due to three proteins H
1, H
2, H
3 with molecular weights of 46, 43 and 40 kD, respectively. These proteins were isolated by preparative PAGE. H
1 and H
2 were shown to be stable in SDS and α-2-ME whereas H
3 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. H
3 was demonstrated to be a bifunctional protein that can lyse and agglutinate erythrocytes. At 56°C hemolytic activity of all three proteins was inactivated, but the agglutination activity of H
3 was stable. Intracoelomic injection of erythrocytes reduced the number of hemolysins from three to two. Monospecific antisera were raised against the isolated hemolysins H
1,2 and 3. The use of these antibodies and of carbohydrates as inhibitors of the biological acivity of the molecules demonstrates the close structural relationship of agglutinins and hemolysins in the CF of
E. fetida.