Application of two‐dimensional electrophoresis and matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry for proteomic analysis of the sexually transmitted parasite Trichomonas vaginalis

Abstract

Trichomonas vaginalis is a sexually transmitted protozoan parasite that infects the human urogenital tract causing trichomoniasis, a worldwide disease. In this work, a fresh clinical isolate of T. vaginalis was used for study of the protein expression in this species. Two-dimensional gel electrophoresis (2-DE) and MALDI-TOF/TOF mass spectrometry (MS) were employed to create a reference map of soluble proteins in the pH range 4-7. A set of 116 proteins belonging to functional classes expressed in high and low abundance was identified by peptide mass fingerprinting and tandem MS. These identifications corresponded to 67 different proteins, suggesting that post-translational modifications are common phenomena in T. vaginalis. Identified proteins were classified into 16 groups according to biological processes. Among detected proteins we identified the major enzymes involved in both cytosolic and hydrogenosomal metabolic pathways, as well as putative protein targets for new drug design. In addition, this analysis allows validation of previous gene predictions confirming the expression of 15 hypothetical proteins. Finally, the findings here reported represent the first reference proteome map of T. vaginalis and the first steps towards the description of a comprehensive proteome map of this parasite.

Immunoproteomics of the active degradome to identify biomarkers for Trichomonas vaginalis

“…The differences observed between the theoretical and the experimental MW and p I of the identified proteinases could be due to the nature of CPs and its processing stages, and/or the CPs putative PTMs. These results are consistent with the proteomic data reported by others 41, 43–45.…”

“…It could also be due to PTMs, phosphorylation and glycosylation; which could be the case for spots 7, 14, 15, and 16 identified as TvLEGU‐1. This behavior is consistent with the data previously reported for other proteinases 39, 41, 43–45, supporting that these differences are due to the nature of CPs and its processing.…”

“…It is relevant to mention that only nine of the 48 papain‐like and three of the 10 legumain‐like CP genes of T. vaginalis seem to be expressed among distinct T. vaginalis isolates displaying high and low virulence, or same isolates grown under different culture conditions 20, 37, 40, 41, 43–45, or distinct isolates grown under normal conditions 45, or even distinct forms of sample preparation. The only identified CPs up to now are TvCP1, TvCP2, TvCP3, TvCP4, TvCP4‐like, TvCP12, TvCPT, TvLEGU‐1, one uncharacterized papain‐like, and two uncharacterized legumain‐like CPs 20, 38–41, 43–45.…”

Identification of the cross-reactive and species-specific antigens between Neospora caninum and Toxoplasma gondii tachyzoites by a proteomics approach

“…Furthermore, the availability of a tenfold coverage of the sequence for the T. gondii genome (www.toxodb.org/ToxoDB.shtml), EST sequence data available for N. caninum (http://genome.wustl.edu/data/ est.cgi), and that of other apicomplexan parasites, such as Plasmodium falciparum, Eimeria acervulina, and Eimeria tenella, has given rise to a preliminary analysis of global protein expression in both parasites (Belli et al 2005). The proteomics approach has been found to be useful for analyzing the proteomes of some parasitic organisms, e.g., Plasmodium (Choumet et al 2007), Toxoplasma (Kawase et al 2007), Trichomonas (De Jesus et al 2007), Leishmania (Brobey et al 2006), and Trypanosoma (Foucher et al 2006) species, including their development, evolution, and pathogenicity. The first proteomics analysis in N. caninum was published by Lee et al (2003).…”

Simple Preparation of Green Fluorescent Protein Conjugated with β-Cyclodextrin in a Site Specific Manner

“…The products were directly applied to spectroscopic and matrix-assisted laser desorption/ionization linear time-of-flight mass spectrometry (MALDI-TOF MS) analysis to confirm the conjugation. Though MALDI-TOF MS is a suitable technique to acquire accurate information of fragmented peptides, 19,20 GFP that adopt a β-barrel topology are highly resistant to proteolysis. 21 We further evaluated fluorescent and structural properties on β-CD grafted GFP through such analysis to certify possibilities of parts for a supramolecule.…”