In vivo intravascular biotinylation of Schistosoma bovis adult worms and proteomic analysis of tegumental surface proteins

Abstract

Schistosoma bovis is a blood-dwelling fluke of ruminants that lives for years inside the vasculature of their hosts. The parasite tegument covers the surface of the worms and plays a key role in the host-parasite relationship. The parasite molecules expressed at the tegument surface are potential targets for immune or drug intervention. The purpose of this work was the identification of the proteins expressed in vivo on the surface of the tegument of S. bovis adult worms. To accomplish this we used a method based on in vivo vascular perfusion of mice infected with S. bovis which allowed the labelling of the surface of the worms inside the blood vasculature. The biotinylation of parasite inside blood vessels prevents the handling of worms in vitro and hence possible damage to the tegument that could produce results that would be difficult to interpret. Trypsin digestion of biotinylated proteins and subsequent liquid chromatography and tandem mass spectrometry analysis (LC-MS/MS) resulted in the identification on the S. bovis tegument of 80 parasite proteins and 28 host proteins. The proteins identified were compared with the findings from other proteomic studies of the schistosome surface. The experimental approach used in this work is a reliable method for selective investigation of the surface of the worms and provides valuable information about the exposed protein repertoire of the tegument of S. bovis in the environmental conditions that the parasite faces inside the blood vessels. To identify the proteins expressed on the surface of the tegument of S. bovis adult worms we used a method based on in vivo vascular perfusion, with biotin, of mice infected with S. bovis which allowed the labelling of the surface of the worms inside the blood vasculature. This methodology prevents the handling of worms in vitro and hence possible damage to the tegument that could produce results that would be difficult to interpret. This work is the first in which vascular perfusion has been used to investigate, in vivo, the protein exposed by an intravascular pathogen on its surface to the host, and provides valuable information about the exposed protein repertoire of the tegument of S. bovis in the environmental conditions that the parasite faces inside the blood vessels.