Carbohydrate profiling and protein identification of tegumental and excreted/secreted glycoproteins of adult Schistosoma bovis worms

Abstract

Schistosoma bovis is a parasite of wild and domestic ruminants that is broadly distributed throughout many tropical and temperate regions of the old world. S. bovis causes severe health problems and significant economic losses in livestock, but in contrast to human schistosomes, S. bovis has been little investigated at a molecular level. Since schistosome glycans and glycoproteins can play important roles in the host-parasite interplay, the aims of the present work were: (i) to characterize the glycans expressed by adult S. bovis worms on their excreted/secreted (ES) and tegumental (TG) glycoproteins and (ii) to identify their carrier protein backbones by mass spectrometry. Using a panel of lectins and monoclonal and polyclonal anti-glycan antibodies, we observed: (i) the absence of sialic acid in S. bovis; (ii) the presence of complex-type N-glycans and LDN antennae on ES glycoproteins; (iii) the presence of glycans containing the Fucalpha1-2Galbeta motif in many TG glycoproteins, and (iv) the presence of glycans containing the Fucalpha1-3GlcNAc motif on many ES and TG glycoproteins but, simultaneously, the absence of the F-LDN(-F) glycans from both the ES and TG glycoproteins. Interestingly, we also found the Lewis(X) and Lewis(Y) antigens co-expressed on several TG isoforms of ATP:guanidino kinase and glyceraldehyde-3-phosphate dehydrogenase. Finally, by ELISA we observed the presence of antibodies against Lewis(X), Lewis(Y) and F-LDN(-F) in the sera of sheep experimentally infected with S. bovis.

Identification of Antigenic Glycans from Schistosoma mansoni by Using a Shotgun Egg Glycan Microarray

“…These global profiling studies have confirmed the major glycan structures previously identified and provided new information on the diversity of glycans and the heterogeneity found among the life stages of the parasite and their secretions (27,(29)(30)(31). Several studies also have demonstrated the feasibility of profiling schistosome glycans using various microarray printing technologies and probing with glycan-binding proteins, antibodies, and antisera, demonstrating that such techniques can be powerful tools for profiling the immune response to this disease (26,(32)(33)(34) and the development or characterization of novel anti-glycan reagents.…”

Identification of antigenic proteins from Echinostoma caproni (Trematoda) recognized by mouse immunoglobulins M, A and G using an immunoproteomic approach

“…After N‐glycosidase digestion and subsequent Western immunoblotting of the ESP in the MW range 70–75 kDa, the enolase was detected at the expected MW. The glycosylation of enolases has been previously reported in other trematodes such as S. bovis (28). In our case the glycosylation of this isoform of E. caproni enolase appears to affect the target epitopes recognized by the antirecombinant enolase antibodies.…”

Echinostomes: genomics and proteomics

“…Our group has obtained preliminary data about the glycosylation state of specific proteins like enolase (Sotillo et al 2008) . The combination of 1DE or 2DE with lectin-binding and mass spectrometry could help to identify and characterize most of the echinostome proteins capable of binding lectins, similar to that already described for Schistosoma bovis (Ramajo-Hernández et al 2007a) .…”