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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-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.

SUBMITTER: Hackler AL 

PROVIDER: S-EPMC4574508 | biostudies-literature | 2015 Dec

REPOSITORIES: biostudies-literature

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Characterization of an African trypanosome mutant refractory to lectin-induced death.

Hackler Amber L AL   Qiu Yijian Y   Patrick Stephen L SL   Lee Soo Hee SH   Acosta-Serrano Alvaro A   Morris James C JC  

Biochemistry and biophysics reports 20151201


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 <i>Trypanosoma brucei</i> 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 compa  ...[more]

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