| Autor: |
Hackler AL; Eukaryotic Pathogens Innovation Center, Department of Genetics and Biochemistry, Clemson University., Qiu Y; Eukaryotic Pathogens Innovation Center, Department of Genetics and Biochemistry, Clemson University., Patrick SL; Eukaryotic Pathogens Innovation Center, Department of Genetics and Biochemistry, Clemson University., Lee SH; Computational Biology and Bioinformatics Organization, Broad Institute, Harvard University and the Massachusetts Institute of Technology., Acosta-Serrano A; Department of Parasitology and Department of Vector Biology, Liverpool School of Tropical Medicine, Liverpool, UK., Morris JC; Eukaryotic Pathogens Innovation Center, Department of Genetics and Biochemistry, Clemson University. |
| Abstrakt: |
Incubation of African trypanosomes with the lectin concanavalin A (conA) leads to alteration in cellular DNA content, DNA degradation, and surface membrane blebbing. Here, we report the generation and characterization of a conA-refractory Trypanosoma brucei line. These insect stage parasites were resistant to conA killing, with a mediun lethal dose at least 50-fold greater than the parental line. Fluorescence-based experiments revealed that the resistant cells bound less lectin when compared to the parental line. Western blotting and mass spectrometry confirmed that the resistant line lacked an N-glycan required for conA binding on the cellular receptors, EP procyclin proteins. The failure to N-glycosylate the EP procyclins was not the consequence of altered N-glycan precursor biosynthesis, as another glycosylated protein (Fla1p) was normally modified. These findings support the likelihood that resistance to conA was a consequence of failure to bind the lectin trigger. |
