Molecular Characterization of Tc964, A Novel Antigenic Protein from Trypanosoma cruzi

• Published in: International journal of molecular sciences Vol. 21; no. 7; p. 2432
• Main Authors: Ruiz-Márvez, Elizabeth, Ramírez, César Augusto, Rodríguez, Eliana Rocío, Flórez, Magda Mellisa, Delgado, Gabriela, Guzmán, Fanny, Gómez-Puertas, Paulino, Requena, José María, Puerta, Concepción J
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 31-03-2020; MDPI
• Subjects: Amino acids; Animals; Antibodies, Protozoan; Antigenicity; Antigens; Antigens, Protozoan - chemistry; Antigens, Protozoan - genetics; Chagas disease; Chagas Disease - diagnosis; Chagas Disease - immunology; Cross Reactions; Cross-reactivity; Cutaneous leishmaniasis; Dynamic structural analysis; Enzymes; Epitopes; Epitopes, B-Lymphocyte; Fractionation; Gene expression; Genomes; GTP Phosphohydrolases; Homology; Humans; Infections; Leishmania infantum - genetics; Leishmania infantum - metabolism; Leishmania major; leishmaniasis; Leishmaniasis - immunology; Leishmaniasis, Cutaneous - immunology; Leishmaniasis, Cutaneous - parasitology; Lymphocytes B; Mass spectrometry; Molecular dynamics; Molecular Dynamics Simulation; molecular modelling; Parasites; Parasitic diseases; Peptides; Phylogeny; Proteins; Protozoa; Protozoan Proteins - chemistry; Protozoan Proteins - genetics; Scientific imaging; serodiagnosis; Serologic Tests; Simulation; Synthetic peptides; Tropical diseases; Trypanosoma cruzi; Trypanosoma cruzi - genetics; Trypanosoma cruzi - metabolism; Virulence; Virulence factors; Latin America; serodiagnosis; chagas disease; leishmaniasis; molecular modelling; Leishmania major; Trypanosoma cruzi
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms21072432

The Tc964 protein was initially identified by its presence in the interactome associated with the LYT1 mRNAs, which code for a virulence factor of
. Tc964 is annotated in the
genome as a hypothetical protein. According to phylogenetic analysis, the protein is conserved in the different genera of the Trypanosomatidae family; however, recognizable orthologues were not identified in other groups of organisms. Therefore, as a first step, an in-depth molecular characterization of the Tc946 protein was carried out. Based on structural predictions and molecular dynamics studies, the Tc964 protein would belong to a particular class of GTPases. Subcellular fractionation analysis indicated that Tc964 is a nucleocytoplasmic protein. Additionally, the protein was expressed as a recombinant protein in order to analyze its antigenicity with sera from Chagas disease (CD) patients. Tc964 was found to be antigenic, and B-cell epitopes were mapped by the use of synthetic peptides. In parallel, the
homologue (Lm964) was also expressed as recombinant protein and used for a preliminary evaluation of antigen cross-reactivity in CD patients. Interestingly, Tc964 was recognized by sera from Chronic CD (CCD) patients at different stages of disease severity, but no reactivity against this protein was observed when sera from Colombian patients with cutaneous leishmaniasis were analyzed. Therefore, Tc964 would be adequate for CD diagnosis in areas where both infections (CD and leishmaniasis) coexist, even though additional assays using larger collections of sera are needed in order to confirm its usefulness for differential serodiagnosis.