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Molecular & Cellular Proteomics 6:346-355, 2007.
© 2007 by The American Society for Biochemistry and Molecular Biology, Inc.

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Research

Proteomics Analysis and Protein Expression during Sporozoite Excystation of Cryptosporidium parvum (Coccidia, Apicomplexa)^*,S

William J. Snelling^,, Qishan Lin^¶, John E. Moore^||, B. Cherie Millar^||, Fabio Tosini^**,, Edoardo Pozio^**,, James S. G. Dooley and Colm J. Lowery^,

From the Centre for Molecular Biosciences, School of Biomedical Sciences, University of Ulster, Cromore Road, Coleraine, County Londonderry BT52 1SA, Northern Ireland, ^¶ UAlbany Proteomics Facility, Centre for Functional Genomics, University of Albany, Rensselaer, New York 12144, ^|| Northern Ireland Public Health Laboratory, Department of Bacteriology, Belfast City Hospital, Belfast BT9 7AD, Northern Ireland, and ^** Department of Infectious, Parasitic, and Immunomediated Diseases, Istituto Superiore di Sanità, Rome 00161, Italy

Cryptosporidiosis, caused by coccidian parasites of the genus Cryptosporidium, is a major cause of human gastrointestinal infections and poses a significant health risk especially to immunocompromised patients. Despite intensive efforts for more than 20 years, there is currently no effective drug treatment against these protozoa. This study examined the zoonotic species Cryptosporidium parvum at two important stages of its life cycle: the non-excysted (transmissive) and excysted (infective) forms. To increase our understanding of the molecular basis of sporozoite excystation, LC-MS/MS coupling with a stable isotope N-terminal labeling strategy using iTRAQ^TM reagents was used on soluble fractions of both non-excysted and excysted sporozoites, i.e. sporozoites both inside and outside oocysts were examined. Sporozoites are the infective stage that penetrates small intestinal enterocytes. Also to increase our knowledge of the C. parvum proteome, shotgun sequencing was performed on insoluble fractions from both non-excysted and excysted sporozoites. In total 303 C. parvum proteins were identified, 56 of which, hitherto described as being only hypothetical proteins, are expressed in both excysted and non-excysted sporozoites. Importantly we demonstrated that the expression of 26 proteins increases significantly during excystation. These excystation-induced proteins included ribosomal proteins, metabolic enzymes, and heat shock proteins. Interestingly three Apicomplexa-specific proteins and five Cryptosporidium-specific proteins augmented in excysted invasive sporozoites. These eight proteins represent promising targets for developing vaccines or chemotherapies that could block parasite entry into host cells.

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