Towards novel therapeutic interventions against helminth infections:structure-function study of P43 in parasitic nematodes using glycoanalytical tools

Lead Research Organisation: University of Manchester
Department Name: Chemistry

Abstract

Soil-transmitted helminth infections affect the poorest and most deprived communities worldwide. They are transmitted by eggs through soil contamination in areas where sanitation is poor. More than 1.5 billion people are affected by soil-transmitted helminth infections. Infections are widely distributed in tropical and subtropical areas, with the greatest numbers occurring in sub-Saharan Africa, the Americas, China and East Asia. Helminth infections are not necessarily lethal, but cause nutritional impairment which is recognized to have a significant impact on growth and physical development, particularly in children ('Soil-transmitted helminth infections', factsheet, WHO 2016). This project aims to investigate the underlying mechanisms by which parasitic nematodes colonise humans.
Previous work by the Grencis group suggests that a 43 kDa protein (P43) is the most dominant secreted molecule from the whipworm (Trichuris sp.). Although the Grencis group has obtained a crystal structure of the protein, its function remains unknown, particularly since the protein does appear to have little similarity with any other protein in databases. Preliminary studies suggest that glycosylation and glycan binding will be important to its function. An understanding of the structure-function of P43 will lead to development of better diagnostic and therapeutic strategies to combat the serious health effects of this infectious disease. This studentship will investigate the
structure of P43-glycans and binding of P43 to glycan arrays using analytical tools (chromatography and mass spectrometry). Understanding the role of glycosylation in P43 will be important for developing P43 as a target for novel therapeutic interventions against helminth infections. The deliverables of this studentship are:
1 Enzyme production. Production of P43 will follow previously established protocols.
2 Structural analysis of glycosylation pattern. Sequence analysis suggests that there are three N-glycosylation sites. A fluorescence and mass spectrometry-active labelling reagent will be used to rapidly prepare released N-glycans for HILIC (hydrophilic interaction liquid chromatography) analysis.
3 Structural sequence identification of N-glycans. The carbohydrate linkage will be sequence identified by IM-MS (Ion Mobility Mass Spectrometry) on industrial placement at Bio-Shape.
4 Structure-function studies: Preliminary studies have suggested that P43 binds to glycans. Glycan array technology compatible with MALDI-ToF MS/MS (matrix-assisted laser desorption/ionization - time of flight) will be used to explore carbohydrate binding.
5 Training and Development. The student will gain experience in a truly transdisciplinary academic-industrial environment being trained in a number of generic techniques including: biological chemistry, enzymology, molecular biology and spectroscopy and spectrometry. Bio-Shape will provide commercial training in the "business model canvass" and exploitation of results.

Publications

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Studentship Projects

Project Reference Relationship Related To Start End Student Name
BB/R505869/1 01/10/2017 30/09/2021
1924636 Studentship BB/R505869/1 01/10/2017 30/09/2018 Hasina Mahmood