Purification of an agglutinin from the haemolymph of the snail Bulinus nasutus and demonstration of related proteins in other Bulinus spp.

Abstract

The snail Bulinus nasutus 1214 possesses a potent haemagglutinin (end-point titre with human erythrocytes, 2(-18)) in its cell-free haemolymph which also binds to the miracidia (but not other larvae) of the incompatible parasite Schistosoma margrebowiei. We have purified a protein possessing this haemagglutinating property from the plasma of this snail. The native Mr of this protein was estimated by SDS polyacrylamide gel electrophoresis to be 210 kDa; under denaturing conditions in a 7.5% PAGE gel it ran as a major band of 135 kDa. Proteins of similar Mr were also found in the haemolymph of 16 other Bulinus spp. (the major intermediate hosts of human and veterinary schistosomiasis in Africa) although the plasma of none of these agglutinated human erythrocytes. Nonetheless, Cleveland mapping of the Mr 135 kDa bands from these different Bulinus spp. revealed 4 identical major peptide fragments (30, 28, 19 and 16 kDa) in each, thus demonstrating a similarity in the primary structure of these plasma proteins. Antisera from Balb/C mice immunized with the 135 kDa polypeptide from Bulinus truncatus 1521 cross-reacted in Western blots with the 135 kDa band of other members of the same truncatus/tropicus species complex but not with species from the africanus or forskalii species groups.