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Originally published In Press as doi:10.1074/mcp.M500287-MCP200 on November 3, 2005.
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Molecular & Cellular Proteomics 5:347-356, 2006.
© 2006 by The American Society for Biochemistry and Molecular Biology, Inc.


Research

Proteins Exposed at the Adult Schistosome Surface Revealed by Biotinylation*,S

Simon Braschi{ddagger} and R. Alan Wilson

From the Department of Biology, University of York, York YO10 5DD, United Kingdom

The human blood-dwelling parasite Schistosoma mansoni can survive in the hostile host environment for decades and must therefore display effective strategies to evade the host immune responses. The surface of the adult worm is covered by a living syncytial layer, the tegument, bounded by a complex multilaminate surface. This comprises a normal plasma membrane overlain by a secreted bilayer, the membranocalyx. Recent proteomic studies have identified constituents of the tegument, but their relative locations remain to be established. We labeled the most exposed surface proteins using two impermeant biotinylation reagents that differed only in length. We anticipated that the two reagents would display distinct powers of penetration, thereby producing a differential labeling pattern. The labeled proteins were recovered by streptavidin affinity and identified by tandem mass spectrometry. A total of 28 proteins was identified, 13 labeled by a long form reagent and the same 13 plus a further 15 labeled by a short form reagent. The parasite proteins included membrane enzymes, transporters, and structural proteins. The short form reagent additionally labeled some cytosolic and cytoskeletal proteins, the latter being constituents of the intracellular spines. Only a single secreted protein was labeled, implying a location between the plasma membrane and the membranocalyx or as part of the latter. Four host proteins, three immunoglobulin heavy chains and C3c/C3dg, a fragment of complement C3, were labeled by both reagents indicating their exposed situation. The presence of the degraded complement C3 implicates inhibition of the classical pathway as a major element of the immune evasion strategy, whereas the recovery of only one truly secreted protein points to the membranocalyx acting primarily as an inert protective barrier between the immune system and the tegument plasma membrane. Collectively the labeled parasite proteins merit investigation as potential vaccine candidates.


{ddagger} To whom correspondence should be addressed: Dept. of Biology, University of York, York YO10 5DD, UK. Tel.: 44-1904-328592; Fax.: 44-1904-328599; E-mail: sb212{at}york.ac.uk


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