Proteomic analysis of the Trypanosoma cruzi ribosomal proteins

Ayub, Maximiliano Juri; Atwood, James; Nuccio, Arthur; Tarleton, Rick L.; Levìn, Mariano J.

doi:10.1016/j.bbrc.2009.02.095

Cited by 22 publications

(24 citation statements)

References 21 publications

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“…Seventy-four are annotated as “hypothetical” in the T. cruzi genome. Seventy five (38 “hypothetical”) were not represented in proteomic data available on TriTrypdb.org (downloaded 4/10/2009) or the ribosomal proteome [14]. One hundred nine were not previously identified in epimastigote data from these sources.…”

Section: Resultsmentioning

confidence: 99%

Identification of Contractile Vacuole Proteins in Trypanosoma cruzi

et al. 2011

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Contractile vacuole complexes are critical components of cell volume regulation and have been shown to have other functional roles in several free-living protists. However, very little is known about the functions of the contractile vacuole complex of the parasite Trypanosoma cruzi, the etiologic agent of Chagas disease, other than a role in osmoregulation. Identification of the protein composition of these organelles is important for understanding their physiological roles. We applied a combined proteomic and bioinfomatic approach to identify proteins localized to the contractile vacuole. Proteomic analysis of a T. cruzi fraction enriched for contractile vacuoles and analyzed by one-dimensional gel electrophoresis and LC-MS/MS resulted in the addition of 109 newly detected proteins to the group of expressed proteins of epimastigotes. We also identified different peptides that map to at least 39 members of the dispersed gene family 1 (DGF-1) providing evidence that many members of this family are simultaneously expressed in epimastigotes. Of the proteins present in the fraction we selected several homologues with known localizations in contractile vacuoles of other organisms and others that we expected to be present in these vacuoles on the basis of their potential roles. We determined the localization of each by expression as GFP-fusion proteins or with specific antibodies. Six of these putative proteins (Rab11, Rab32, AP180, ATPase subunit B, VAMP1, and phosphate transporter) predominantly localized to the vacuole bladder. TcSNARE2.1, TcSNARE2.2, and calmodulin localized to the spongiome. Calmodulin was also cytosolic. Our results demonstrate the utility of combining subcellular fractionation, proteomic analysis, and bioinformatic approaches for localization of organellar proteins that are difficult to detect with whole cell methodologies. The CV localization of the proteins investigated revealed potential novel roles of these organelles in phosphate metabolism and provided information on the potential participation of adaptor protein complexes in their biogenesis.

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Section: Resultsmentioning

confidence: 99%

Identification of Contractile Vacuole Proteins in Trypanosoma cruzi

et al. 2011

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“…These extensions are only present in kinetoplastids, although their length varies among species. MS analyses of T. cruzi ribosomes confirmed the presence of peptides matching to TcL19 and TcS21, strongly suggesting that these genes correspond to the functional ribosomal components [6]. The possible functional roles of these extensions, as well as the molecular mechanisms that generated them over time, constitute interesting fields for future studies.…”

Section: S Cerevisiaementioning

confidence: 83%

“…However, the presence of specific features due to ribosomal proteins is difficult to demonstrate by this technique. Therefore, using the S. cerevisiae ribosomal protein sequences as probes, it was possible to identify in the T. cruzi genome database all homologue genes [6]. The average amino acid identity between the S. cerevisiae and T. cruzi ribosomal proteins was remarkably low (~50%), taking into account the high degree of conservation of the ribosome through evolution.…”

Section: Sequence and Proteomic Analysis: Differences On The Ribosomamentioning

confidence: 99%

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Ribosomes from Trypanosomatids: Unique Structural and Functional Properties

Ayub¹,

Lapadula²,

Hoebeke³

et al. 2012

Cell-Free Protein Synthesis

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Additional information is available at the end of the chapter http://dx.doi.org/10.5772/48336 IntroductionTrypanosomatids are a monophyletic group of protozoa that diverged early from the eukaryotic lineage, constituting valuable model organisms for studying variability in different highly conserved processes including protein synthesis. Moreover, several species of trypanosomatids are causing agents of endemic diseases in the third world. There are many evidences suggesting that translation in these organisms shows important differences with that of model organisms such as yeast and mammals. These unique features, which have a great potential relevance for both basic and applied research, will be discussed in this chapter. Structural analysis Cryo-electron microscopy map of Trypanosoma cruzi ribosome: Unique features of the rRNAUsing the cryo-electron microscopy (cryo-EM) technique, a 12Å resolution density map of the T. cruzi 80S ribosome has been constructed [1]. The overall structure of the T. cruzi 80S ribosome exhibits well defined small (40S) and large (60S) subunits (Figure 1). Some of the landmark characteristics of the ribosome structure can be identified in the density map. Compared with the 80S ribosome from yeast, both the small and large ribosomal subunits from T. cruzi are larger, mainly due to the size of the ribosomal RNA molecules. T. cruzi rRNA (18S rRNA: 2,315 nt and 28S rRNA: 4,151 nt) is one-fifth larger than yeast rRNA (18S rRNA: 1,798 nt; 25S rRNA: 3,392 nt) in total number of nucleotides.Although the T. cruzi 80S ribosome possesses conserved ribosomal structures, it exhibits many distinctive structural features in both the small and large subunits. Compared with other eukaryotic ribosomes, the T. cruzi ribosomal 40S subunit appears expanded, due to the addition of a large piece of density adjacent to the platform region ( Figure 1). As can be seen in the secondary structure of the T. cruzi 18S rRNA (Figure 2), this extra density must be attributed to two large expansion segments (ES) in domain II of the 18S rRNA, ES6 and ES7, designated as insertions of helices 21 and 26. These are the two largest ES in the T. cruzi 18S rRNA, involving 504 and 147 nucleotides, respectively.

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“…Repeated motifs are found in several members of the 60S ribosomal proteins [19]. The largest C terminal extensions (more than 160 amino acids) have been observed in T. cruzi L19 and T. cruzi S21 and are specific to trypanosomatids [20] …”

Section: Introductionmentioning

confidence: 99%

Anti-Trypanosoma cruzi Cross-Reactive Antibodies Detected at High Rate in Non-Exposed Individuals Living in Non-Endemic Regions: Seroprevalence and Association to Other Viral Serologies

Saba

Gueyffier²,

Dichtel-Danjoy³

et al. 2013

PLoS ONE

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Cross-reactive antibodies are characterized by their recognition of antigens that are different from the trigger immunogen. This happens when the similarity between two different antigenic determinants becomes adequate enough to enable a specific binding with such cross-reactive antibodies. In the present manuscript, we report the presence, at an “abnormal” high frequency, of antibodies in blood samples from French human subjects cross-reacting with a synthetic-peptide antigen derived from a Trypanosoma cruzi (T. cruzi) protein sequence. As the vector of T. cruzi is virtually confined to South America, the parasite is unlikely to be the trigger immunogen of the cross-reactive antibodies detected in France. At present, the cross-reactive antibodies are measured by using an in-house ELISA method that employs the T. cruzi -peptide antigen. However, to underline their cross-reactive characteristics, we called these antibodies “Trypanosoma cruzi Cross Reactive Antibodies” or TcCRA. To validate their cross-reactive nature, these antibodies were affinity-purified from plasma of healthy blood donor and were then shown to specifically react with the T. cruzi parasite by immunofluorescence. Seroprevalence of TcCRA was estimated at 45% in serum samples of French blood donors while the same peptide-antigen reacts with about 96% of T. cruzi -infected Brazilian individuals. In addition, we compared the serology of TcCRA to other serologies such as HSV 1/2, EBV, HHV-6, CMV, VZV, adenovirus, parvovirus B19, mumps virus, rubella virus, respiratory syncytial virus, measles and enterovirus. No association was identified to any of the tested viruses. Furthermore, we tested sera from different age groups for TcCRA and found a progressive acquisition starting from early childhood. Our findings show a large seroprevalence of cross-reactive antibodies to a well-defined T. cruzi antigen and suggest they are induced by a widely spread immunogen, acquired from childhood. The etiology of TcCRA and their clinical relevance still need to be investigated.

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Proteomic analysis of the Trypanosoma cruzi ribosomal proteins

Cited by 22 publications

References 21 publications

Identification of Contractile Vacuole Proteins in Trypanosoma cruzi

Identification of Contractile Vacuole Proteins in Trypanosoma cruzi

Ribosomes from Trypanosomatids: Unique Structural and Functional Properties

Anti-Trypanosoma cruzi Cross-Reactive Antibodies Detected at High Rate in Non-Exposed Individuals Living in Non-Endemic Regions: Seroprevalence and Association to Other Viral Serologies

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