1. In-solution behavior and protective potential of asparagine synthetase A from Trypanosoma cruzi.
- Author
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Souza Morini F, de Castro E, de Morais SB, Lancheros Contreras C, Pereira Weiler AV, Murakami MT, Pinge-Filho P, Yamada-Ogatta SF, and de Arruda Campos Brasil de Souza T
- Subjects
- Ammonia metabolism, Animals, Aspartate-Ammonia Ligase chemistry, Aspartate-Ammonia Ligase isolation & purification, Chagas Disease prevention & control, Chromatography, Liquid, Circular Dichroism, Disease Models, Animal, Dynamic Light Scattering, Glutamine metabolism, Mice, Inbred BALB C, Parasitemia prevention & control, Protein Conformation, Protein Folding, Protozoan Vaccines administration & dosage, Recombinant Proteins chemistry, Recombinant Proteins isolation & purification, Vaccines, Synthetic administration & dosage, Asparagine metabolism, Aspartate-Ammonia Ligase immunology, Aspartate-Ammonia Ligase metabolism, Recombinant Proteins immunology, Recombinant Proteins metabolism, Trypanosoma cruzi enzymology
- Abstract
l-Asparagine synthetase (AS) acts in asparagine formation and can be classified into two families: AS-A or AS-B. AS-A is mainly found in prokaryotes and can synthetize asparagine from ammonia. Distinct from other eukaryotes, Trypanosoma cruzi produces an AS-A. AS-A from Trypanosoma cruzi (Tc-AS-A) differs from prokaryotic AS-A due to its ability to catalyze asparagine synthesis using both glutamine and ammonia as nitrogen sources. Regarding these peculiarities, this work uses several biophysical techniques to provide data concerning the Tc-AS-A in-solution behavior. Tc-AS-A was produced as a recombinant and purified by three chromatography steps. Circular dichroism, dynamic light scattering, and analytical size exclusion chromatography showed that Tc-AS-A has the same fold and quaternary arrangement of prokaryotic AS-A. Despite the tendency of protein to aggregate, stable dimers were obtained when solubilization occurred at pH ≤ 7.0. We also demonstrate the protective efficacy against T. cruzi infection in mice immunized with Tc-AS-A. Our results indicate that immunization with Tc-AS-A might confer partial protection to infective forms of T. cruzi in this particular model., (Copyright © 2019 Elsevier B.V. All rights reserved.)
- Published
- 2019
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