Proteomic analysis of the schistosome tegument and its surface membranes.
The tegument surface of the adult schistosome, bounded by a normal plasma membrane overlain by a secreted membranocalyx, holds the key to understanding how schistosomes evade host immune responses. Recent advances in mass spectrometry (MS), and the sequencing of the Schistosoma mansoni transcriptome/genome, have facilitated schistosome proteomics. We detached the tegument from the worm body and enriched its surface membranes by differential extraction, before subjecting the preparation to liquid chromatography-based proteomics to identify its constituents. The most exposed proteins on live worms were labelled with impearmeant biotinylation reagents, and we also developed methods to isolate the membranocalyx for analysis. We identified transporters for sugars, amino acids, inorganic ions and water, which confirm the importance of the tegument plasma membrane in nutrient acquisition and solute balance. Enzymes, including phosphohydrolases, esterases and carbonic anhydrase were located with their catalytic domains external to the plasma membrane, while five tetraspanins, annexin and dysferlin were implicated in membrane architecture. In contrast, few parasite proteins could be assigned to the membranocalyx but mouse immune response proteins, including three immunoglobulins and two complement factors, were detected, plus host membrane proteins such as CD44, integrin and a complement regulatory protein, testifying to the acquisitive properties of the secreted bilayer.
Braschi
Simon
S
Department of Biology, University of York, York, United Kingdom. simon.braschi@googlemail.com
Borges
William Castro
WC
Wilson
R Alan
RA
D000818:Animals; D015180:Electrophoresis, Gel, Two-Dimensional; D013058:Mass Spectrometry; D008565:Membrane Proteins / Q000032:analysis* / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data; D020543:Proteome / Q000032:analysis* / Q000235:genetics; D015800:Protozoan Proteins / Q000032:analysis* / Q000235:genetics / Q000302:isolation & purification; D012550:Schistosoma mansoni / Q000737:chemistry* / Q000235:genetics
D008565:Membrane Proteins; D020543:Proteome; D015800:Protozoan Proteins