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Molecular & Cellular Proteomics 5:835-844, 2006.
© 2006 by The American Society for Biochemistry and Molecular Biology, Inc.

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Research

Identification of Novel Proteases and Immunomodulators in the Secretions of Schistosome Cercariae That Facilitate Host Entry ^*,S

Rachel S. Curwen, Peter D. Ashton, Shobana Sundaralingam and R. Alan Wilson

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

Schistosomiasis, caused by parasitic helminths, remains a serious human disease in the tropics. Cercariae of Schistosoma mansoni infect their hosts by direct skin penetration, aided by secretions from acetabular and head glands. Both proteolytic and immunomodulatory properties have been ascribed to the released material, but to date only five isoforms of elastase and one putative anti-inflammatory protein (Sm16) have been cloned. We analyzed secretions from mechanically transformed cercariae by two-dimensional electrophoresis. An average gel image was created and compared with a separation of soluble larval extract, revealing a less complex spot pattern in the secretions with 60% of the spots matched to the larval extract. Subsequent tandem mass spectrometric analysis identified 48 spots from the released material, representing approximately 80% of its normalized volume. Twenty-nine of these are likely to originate in the vesicles, and 18 are likely to originate in the cytosol of the glands (the latter class being present due to holocrine secretion); one is unknown. The vesicular proteins were significantly more enriched than the cytosolic proteins in the released material when compared with the larval extract. A novel metalloproteinase (termed SmPepM8) was the second most abundant constituent after three isoforms of cercarial elastase. In addition, a dipeptidyl peptidase IV (SmDPP IV) was discovered but in much smaller quantity. A new serine protease inhibitor (SmSerp_c) was also prominent. Along with Sm16, four potential immunomodulators were identified, three with similarity to venom allergens (SmSCP_a, _b, and _c) and one with homology to the potassium channel blockers in scorpion venom (SmKK7). Interrogation of the expressed sequence tag database found transcripts encoding the majority of vesicular proteins present solely in the intramolluscan stages of the life cycle. Distinct patterns of radiolabel incorporation suggested three separate origins for the vesicular proteins. All the novel constituents merit investigation as vaccine candidates, and the potential immunomodulators merit investigation as therapeutic agents.

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