Comparative analyses of venoms from American and African Sicarius spiders that differ in sphingomyelinase D activity

Spider venoms are cocktails of toxic proteins and peptides, whose composition varies at many levels. Understanding patterns of variation in chemistry and bioactivity is fundamental for understanding factors influencing variation. The venom toxin sphingomyelinase D (SMase D) in sicariid spider venom (Loxosceles and Sicarius) causes dermonecrotic lesions in mammals. Multiple forms of venom-expressed genes with homology to SMase D are expressed in venoms of both genera. SMase D activity levels differ among major clades with American Sicarius vastly reduced relative to all Loxosceles and African Sicarius despite expression of SMase D homologs in venoms of American Sicarius. Here we report comparative analyses of protein composition and insecticidal activity of crude venoms from three Sicarius species, two from South Africa and one from Central America. Comparative 2-dimensional electrophoresis shows dense regions of proteins in the size range of SMase D in all three species, but there are differences in sizes and isoelectric points (pIs). Few proteins strictly co-migrate and there are clusters of proteins with similar pIs and molecular weights whose patterns of similarity do not necessarily reflect phylogenetic relatedness. In addition, PD50 estimates on crickets indicate a small though significant decrease in potency of South American Sicarius venoms relative to African species.