This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Trypanosoma cruzi (T. cruzi) is a protozoan parasite endemic to Latin America and the causative agent of Chagas' disease. The life cycle of T. cruzi is complex, with multiple developmental stages persisting between a variety of mammalian host (including humans) and insect vectors. Although the cell surface glycoproteins of T. cruzi have been implied in numerous critical functions, such as host cell recognition, host cell invasion, and immune evasion, little is actually known on this topic. We are developing tools to analyze the Glycoproteome to provide information on this subject. In this strategy, tryptic glycopeptides are isolated using serial lectin affinity chromatography. The pool of glycopeptides eluted from each lectin column is then subjected to enzymatic de-glycosylation which incorporates an 18O into the occupied glycosylation sites permitting their identification by MS/MS. The structures of the released glycans are analyzed by MALDI-MS after permethylation, while the formerly glycosylated peptides are identified by LC-MS/MS.
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