Quantitative phosphoproteome and proteome analyses emphasize the influence of phosphorylation events during the nutritional stress of Trypanosoma cruzi: the initial moments of in vitro metacyclogenesis

Abstract

Phosphorylation is an important event in cell signaling that is modulated by kinases and phosphatases. In Trypanosoma cruzi, the etiological agent of Chagas disease, approximately 2% of the protein-coding genes encode for protein kinases. This parasite has a heteroxenic life cycle with four different development stages. In the midgut of invertebrate vector, epimastigotes differentiate into metacyclic trypomastigotes in a process known as metacyclogenesis. This process can be reproduced in vitro by submitting parasites to nutritional stress (NS). Aiming to contribute to the elucidation of mechanisms that trigger metacyclogenesis, we applied super-SILAC (super-stable isotope labeling by amino acids in cell culture) and LC-MS/MS to analyze different points during NS. This analysis resulted in the identification of 4205 protein groups and 3643 phosphopeptides with the location of 4846 phosphorylation sites. Several phosphosites were considered modulated along NS and are present in proteins associated with various functions, such as fatty acid synthesis and the regulation of protein expression, reinforcing the importance of phosphorylation and signaling events to the parasite. These modulated sites may be triggers of metacyclogenesis.

Molecular dissection of Chagas induced cardiomyopathy reveals central disease associated and druggable signaling pathways

“…Previous studies applying proteomics to investigate Chagas disease have mostly been focused on serum, looking for disease progression markers using two-dimensional gel electrophoresis prior to mass spectrometry in rats [67] and human patients [68,69]. Also, proteomic and phospho-proteomic approaches have been targeted toward the parasite itself, describing its response to nutritional stress [70] and proteins modulated during the differentiation process [71,72]. However, a large-scale, phospho-proteomic analysis of chagasic hearts, one of the most affected organs, has not been performed.…”

Extensive Translational Regulation through the Proliferative Transition of Trypanosoma cruzi Revealed by Multi-Omics

“…Similar yields were previously reported in trypanosomatid ribosome profiling applying the same cutoff for detection (7,873 transcripts in T. cruzi [ 23 ], 7,792 [ 69 ] and 7,773 [ 70 ] transcripts in T. brucei ). Likewise, recent proteomic studies of T. cruzi detected 4,060 ( 71 ) and 4,205 ( 72 ) protein groups, while a T. brucei cell cycle study detected 4,629 protein groups ( 46 ). Therefore, the results of the two omic approaches we carried out to investigate G 1 /S cell cycle phase transition in T. cruzi are consistent with the current state of the art in trypanosomatids regarding sensitivity and coverage.…”

Proteomic Insights into the Biology of the Most Important Foodborne Parasites in Europe

“…Quantitation of phosphorylated proteins in the same study showed that there are more than 7000 phosphorylation sites, of which 260 are under different regulation, including some potential drug targets, e.g., sterol biosynthesis enzymes. Next study indicated that during periods of nutritional stress, many proteins are phosphorylated, which may trigger metacyclogenesis [ 150 ]. Similarly, the proteomic comparison of the exponential phase and stationary phase of the epimastigotes quantified more than 3000 proteins [ 151 ].…”