Evelyn Rogerson
Variations in the Peritrophic Matrix Composition of Heparan Sulphate from the Tsetse Fly, Glossina morsitans morsitans
Rogerson, Evelyn; Pelletier, Julien; Acosta-Serrano, Alvaro; Rose, Clair; Taylor, Sarah; Guimond, Scott; Lima, Marcelo; Skidmore, Mark; Yates, Edwin
Authors
Julien Pelletier
Alvaro Acosta-Serrano
Clair Rose
Sarah Taylor
Scott Guimond s.e.guimond@keele.ac.uk
Marcelo Andrade De Lima m.andrade.de.lima@keele.ac.uk
Mark Skidmore m.a.skidmore@keele.ac.uk
Edwin Yates
Abstract
Tsetse flies are the principal insect vectors of African trypanosomes-sleeping sickness in humans and Nagana in cattle. One of the tsetse fly species, Glossina morsitans morsitans, is host to the parasite, Trypanosoma brucei, a major cause of African trypanosomiasis. Precise details of the life cycle have yet to be established, but the parasite life cycle involves crossing the insect peritrophic matrix (PM). The PM consists of the polysaccharide chitin, several hundred proteins, and both glycosamino- and galactosaminoglycan (GAG) polysaccharides. Owing to the technical challenges of detecting small amounts of GAG polysaccharides, their conclusive identification and composition have not been possible until now. Following removal of PMs from the insects and the application of heparinases (bacterial lyase enzymes that are specific for heparan sulphate (HS) GAG polysaccharides), dot blots with a HS-specific antibody showed heparan sulphate proteoglycans (HSPGs) to be present, consistent with Glossina morsitans morsitans genome analysis, as well as the likely expression of the HSPGs syndecan and perlecan. Exhaustive HS digestion with heparinases, fluorescent labeling of the resulting disaccharides with BODIPY fluorophore, and separation by strong anion exchange chromatography then demonstrated the presence of HS for the first time and provided the disaccharide composition. There were no significant differences in the type of disaccharide species present between genders or between ages (24 vs. 48 h post emergence), although the HS from female flies was more heavily sulphated overall. Significant differences, which may relate to differences in infection between genders or ages, were evident, however, in overall levels of 2-O-sulphation between sexes and, for females, between 24 and 48 h post-emergence, implying a change in expression or activity for the 2-O-sulphotransferase enzyme. The presence of significant quantities of disaccharides containing the monosaccharide GlcNAc6S contrasts with previous findings in Drosophila melanogaster and suggests subtle differences in HS fine structure between species of the Diptera.
Citation
Rogerson, E., Pelletier, J., Acosta-Serrano, A., Rose, C., Taylor, S., Guimond, S., …Yates, E. (2018). Variations in the Peritrophic Matrix Composition of Heparan Sulphate from the Tsetse Fly, Glossina morsitans morsitans. Pathogens an international, open access journal of pathogens and pathogen-host interactions, 7, https://doi.org/10.3390/pathogens7010032
Journal Article Type | Article |
---|---|
Acceptance Date | Mar 17, 2018 |
Publication Date | Mar 19, 2018 |
Journal | Pathogens |
Print ISSN | 2076-0817 |
Publisher | MDPI |
Volume | 7 |
DOI | https://doi.org/10.3390/pathogens7010032 |
Keywords | Glossina morsitans morsitans, Trypanosoma brucei, heparan sulphate, peritrophic matrix, tsetse fly |
Publisher URL | https://www.ncbi.nlm.nih.gov/pubmed/29562674 |
Files
pathogens-07-00032.pdf
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Publisher Licence URL
https://creativecommons.org/licenses/by/4.0/
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