Molecular Characterization of Tc964, A Novel Antigenic Protein from Trypanosoma cruzi
Abstract
:1. Introduction
2. Results
2.1. Tc964 Is Conserved in Different T. cruzi Discrete Typing Units (DTU) and Is Also Present in Other Kinetoplastids
2.2. Tc964 Protein May Be a Member of the GTPase Family
2.3. Tc964 is a Nucleocytoplasmic Protein
2.4. Tc964 is Recognized by Sera from Chronic Chagas Disease Patients
2.5. Tc964 Protein is Not Recognized by Sera from Leishmaniasis Patients
2.6. Two Peptides Derived from Tc964 Protein are Recognized by Sera from Chronic CD Patients
3. Discussion
4. Materials and Methods
4.1. Description of the TcCLB.511467.70 Gene in the Genome of T. cruzi
4.2. Phylogenetic Analysis of Protein Tc964
4.3. Fractionation and ELISA Test
4.3.1. Cultivation of Parasites
4.3.2. Subcellular Location of the Tc964 Protein of T. cruzi
4.3.3. Soluble Antigens from T. cruzi and L. major
4.4. Western Blot Assay
4.5. Structural Prediction of the Tc964 protein in T. cruzi and L. major
4.6. Cloning and Protein Expression
4.7. Prediction of Linear B-cell Epitopes and Peptide Synthesis
4.8. Sera Samples and Ethical Statement
4.9. ELISA Assay Protocol
4.10. Statistical and Data Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
Abbreviations
ACL | Active Cutaneous Leishmaniasis |
CD | Chagas Disease |
CCD | Chronic Chagas Disease |
CCL | Curated Cutaneous Leishmaniasis |
DTU | Discrete Typing Unit |
ELISA | Enzyme-linked immunosorbent assay |
kDa | Kilodaltons |
PBS | phosphate buffered saline |
RI | Reactivity index |
rLm964 | Recombinant protein Lm964 |
rTc964 | Recombinant protein Tc964 |
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Ruiz-Márvez, E.; Ramírez, C.A.; Rodríguez, E.R.; Flórez, M.M.; Delgado, G.; Guzmán, F.; Gómez-Puertas, P.; Requena, J.M.; Puerta, C.J. Molecular Characterization of Tc964, A Novel Antigenic Protein from Trypanosoma cruzi. Int. J. Mol. Sci. 2020, 21, 2432. https://doi.org/10.3390/ijms21072432
Ruiz-Márvez E, Ramírez CA, Rodríguez ER, Flórez MM, Delgado G, Guzmán F, Gómez-Puertas P, Requena JM, Puerta CJ. Molecular Characterization of Tc964, A Novel Antigenic Protein from Trypanosoma cruzi. International Journal of Molecular Sciences. 2020; 21(7):2432. https://doi.org/10.3390/ijms21072432
Chicago/Turabian StyleRuiz-Márvez, Elizabeth, César Augusto Ramírez, Eliana Rocío Rodríguez, Magda Mellisa Flórez, Gabriela Delgado, Fanny Guzmán, Paulino Gómez-Puertas, José María Requena, and Concepción J. Puerta. 2020. "Molecular Characterization of Tc964, A Novel Antigenic Protein from Trypanosoma cruzi" International Journal of Molecular Sciences 21, no. 7: 2432. https://doi.org/10.3390/ijms21072432