title: Characterization of an African trypanosome mutant refractory to lectin-induced death.
abstract: 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-refractoryTrypanosoma bruceiline. 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.
authors.affiliation: Eukaryotic Pathogens Innovation Center, Department of Genetics and Biochemistry, Clemson University.
authors.forename: Amber L
authors.initials: AL
authors.lastname: Hackler
authors.affiliation: Eukaryotic Pathogens Innovation Center, Department of Genetics and Biochemistry, Clemson University.
authors.forename: Yijian
authors.initials: Y
authors.lastname: Qiu
authors.affiliation: Eukaryotic Pathogens Innovation Center, Department of Genetics and Biochemistry, Clemson University.
authors.forename: Stephen L
authors.initials: SL
authors.lastname: Patrick
authors.affiliation: Computational Biology and Bioinformatics Organization, Broad Institute, Harvard University and the Massachusetts Institute of Technology.
authors.forename: Soo Hee
authors.initials: SH
authors.lastname: Lee
authors.affiliation: Department of Parasitology and Department of Vector Biology, Liverpool School of Tropical Medicine, Liverpool, UK.
authors.forename: Alvaro
authors.initials: A
authors.lastname: Acosta-Serrano
authors.affiliation: Eukaryotic Pathogens Innovation Center, Department of Genetics and Biochemistry, Clemson University.
authors.forename: James C
authors.initials: JC
authors.lastname: Morris
keywords: Trypanosoma brucei; concanavalin A; procyclins; surface molecule