Abstract
The protozoan parasite Trypanosoma brucei has a complex digenetic lifecycle between a mammalian host and an insect vector, and adaption of its proteome between lifecycle stages is essential to its survival and virulence. We have optimized a procedure for growing Trypanosoma brucei procyclic form cells in conditions suitable for stable isotope labeling by amino acids in culture (SILAC) and report a comparative proteomic analysis of cultured procyclic form and bloodstream form T. brucei cells. In total we were able to identify 3959 proteins and quantify SILAC ratios for 3553 proteins with a false discovery rate of 0.01. A large number of proteins (10.6%) are differentially regulated by more the 5-fold between lifecycle stages, including those involved in the parasite surface coat, and in mitochondrial and glycosomal energy metabolism. Our proteomic data is broadly in agreement with transcriptomic studies, but with significantly larger fold changes observed at the protein level than at the mRNA level.
Highlights
Trypanosoma brucei is a protozoan parasite transmitted by the bite of the tsetse fly, and is the etiological agent of African sleeping sickness
T. brucei has a complex digenetic lifecycle between an insect vector and mammalian host, and the ability to respond to its environment through adaption of its proteome is essential to its survival and virulence
The lifecycle is completed by migration to the salivary glands and transformation to an adherent epimastigote form, followed by transformation to a detached metacyclic form, which is competent for transmission into the bloodstream of the mammalian host when the tsetse takes a blood-meal
Summary
Trypanosoma brucei is a protozoan parasite transmitted by the bite of the tsetse fly, and is the etiological agent of African sleeping sickness. T. brucei has a complex digenetic lifecycle between an insect vector and mammalian host, and the ability to respond to its environment through adaption of its proteome is essential to its survival and virulence. The pleomorphic bloodstream form exists as both a replicative long-slender morphology and a division arrested stumpy form which is preadapted for transmission into the tsetse fly. Upon ingestion by the tsetse fly the parasite differentiates into a replicative procyclic form to enable survival in its new environment. The lifecycle is completed by migration to the salivary glands and transformation to an adherent epimastigote form, followed by transformation to a detached metacyclic form, which is competent for transmission into the bloodstream of the mammalian host when the tsetse takes a blood-meal
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