Development of GlycoPhage Arrays as a novel high-t.. (GlycoPhageApp)
Development of GlycoPhage Arrays as a novel high-throughput method for glycomics
Start date: Oct 1, 2012,
End date: Sep 30, 2016
Complex carbohydrates (glycans) are attached to proteins and lipids by the process of glycosylation and play important roles in many biological processes including cell-cell recognition, metabolic trafficking and host-pathogen interactions. Altered glycosylation or variations in the synthesis of glycans are known to cause diseases including cancer, retroviral infection and disorders of the heart, lung and blood. In order to establish connections between glycan structures and their functions (functional glycomics), to monitor glycosylation in disease diagnosis and prognosis, and to elucidate molecular mechanisms involved in pathogenesis, the development of precise, robust and sensitive methodologies for glycan analysis is critical. Carbohydrate-based arrays, or “glycoarrays,” have emerged in the last decade as a powerful tool, however to fully exploit the potential of arrays, it will be necessary to (i) increase the quantity and diversity of carbohydrate structures and (ii) develop reliable and reproducible chemistries for the immobilization of the carbohydrate probes onto solid support. Recently, the protein glycosylation locus (Pgl) discovered in Campylobacter jejuni was functionally transferred to E. coli, conferring ability to glycosylate proteins. Additionally, Dr. Celik has recently demonstrated glycosylation of phage particles simply by infecting the glycosylation competent E. coli with M13 phage displaying an acceptor protein. The hypothesis of this particular application is that the presentation of N-glycosylated proteins and O-antigens on phage particles can be exploited for the development of glycan arrays. The study will be significant because it will overcome the current bottlenecks in glycan array construction and provide a relatively inexpensive, specific and stable glycan representation method, as well as introduce a simplified and universal purification technique that is not dependent on the carbohydrate.
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