The tegument surface membranes of the human blood parasite Schistosoma mansoni: a proteomic analysis after differential extraction.
The blood fluke Schistosoma mansoni can live for years in the hepatic portal system of its human host and so must possess very effective mechanisms of immune evasion. The key to understanding how these operate lies in defining the molecular organisation of the exposed parasite surface. The adult worm is covered by a syncytial tegument, bounded externally by a plasma membrane and overlain by a laminate secretion, the membranocalyx. In order to determine the protein composition of this surface, the membranes were detached using a freeze/thaw technique and enriched by sucrose density gradient centrifugation. The resulting preparation was sequentially extracted with three reagents of increasing solubilising power. The extracts were separated by 2-DE and their protein constituents were identified by MS/MS, yielding predominantly cytosolic, cytoskeletal and membrane-associated proteins, respectively. After extraction, the final pellet containing membrane-spanning proteins was processed by liquid chromatographic techniques before MS. Transporters for sugars, amino acids, ions and other solutes were found together with membrane enzymes and proteins concerned with membrane structure. The proteins identified were categorised by their function and putative location on the basis of their homology with annotated proteins in other organisms.
Braschi
Simon
S
Department of Biology, University of York, York, UK. sb212@york.ac.uk
Curwen
Rachel S
RS
Ashton
Peter D
PD
Verjovski-Almeida
Sergio
S
Wilson
Alan
A
D000818:Animals; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D013058:Mass Spectrometry; D008565:Membrane Proteins / Q000032:analysis* / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data; D020543:Proteome / Q000032:analysis*; D015800:Protozoan Proteins / Q000032:analysis* / Q000235:genetics / Q000302:isolation & purification; D012550:Schistosoma mansoni / Q000033:anatomy & histology / Q000737:chemistry*
D008565:Membrane Proteins; D020543:Proteome; D015800:Protozoan Proteins