Kinetic characterization of a novel cysteine peptidase from the protozoan Babesia bovis, a potential target for drug design
C1A cysteine peptidases have been shown to play an important role during apicomplexan invasion and egress of host red blood cells (RBCs) and therefore have been exploited as targets for drug development, in which peptidase specificity is deterministic. Babesia bovis genome is currently available and...
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| Format: | Artículo |
| Language: | Inglés |
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Elsevier
2022
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| Online Access: | http://hdl.handle.net/20.500.12123/11122 https://www.sciencedirect.com/science/article/abs/pii/S0300908420302182 https://doi.org/10.1016/j.biochi.2020.09.012 |
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| author | Lu, Stephen Ascencio, Mariano Torquato, Ricardo J.S. Florin-Christensen, Mónica Tanaka, Aparecida S. |
| author_browse | Ascencio, Mariano Florin-Christensen, Mónica Lu, Stephen Tanaka, Aparecida S. Torquato, Ricardo J.S. |
| author_facet | Lu, Stephen Ascencio, Mariano Torquato, Ricardo J.S. Florin-Christensen, Mónica Tanaka, Aparecida S. |
| author_sort | Lu, Stephen |
| collection | INTA Digital |
| description | C1A cysteine peptidases have been shown to play an important role during apicomplexan invasion and egress of host red blood cells (RBCs) and therefore have been exploited as targets for drug development, in which peptidase specificity is deterministic. Babesia bovis genome is currently available and from the 17 putative cysteine peptidases annotated four belong to the C1A subfamily. In this study, we describe the biochemical characterization of a C1A cysteine peptidase, named here BbCp (B. bovis cysteine peptidase) and evaluate its possible participation in the parasite asexual cycle in host RBCs. The recombinant protein was obtained in bacterial inclusion bodies and after a refolding process, presented typical kinetic features of the cysteine peptidase family, enhanced activity in the presence of a reducing agent, optimum pH between 6.5 and 7.0 and was inhibited by cystatins from R. microplus. Moreover, rBbCp substrate specificity evaluation using a peptide phage display library showed a preference for Val > Leu > Phe. Finally, antibodies anti-rBbCp were able to interfere with B. bovis growth in vitro, which highlights the BbCp as a potential target for drug design. |
| format | Artículo |
| id | INTA11122 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2022 |
| publishDateRange | 2022 |
| publishDateSort | 2022 |
| publisher | Elsevier |
| publisherStr | Elsevier |
| record_format | dspace |
| spelling | INTA111222022-01-14T11:49:29Z Kinetic characterization of a novel cysteine peptidase from the protozoan Babesia bovis, a potential target for drug design Lu, Stephen Ascencio, Mariano Torquato, Ricardo J.S. Florin-Christensen, Mónica Tanaka, Aparecida S. Cisteína Babesia bovis Peptidasas Proteínas Recombinantes Anticuerpos Cysteine Peptidases Recombinant Proteins Antibodies C1A cysteine peptidases have been shown to play an important role during apicomplexan invasion and egress of host red blood cells (RBCs) and therefore have been exploited as targets for drug development, in which peptidase specificity is deterministic. Babesia bovis genome is currently available and from the 17 putative cysteine peptidases annotated four belong to the C1A subfamily. In this study, we describe the biochemical characterization of a C1A cysteine peptidase, named here BbCp (B. bovis cysteine peptidase) and evaluate its possible participation in the parasite asexual cycle in host RBCs. The recombinant protein was obtained in bacterial inclusion bodies and after a refolding process, presented typical kinetic features of the cysteine peptidase family, enhanced activity in the presence of a reducing agent, optimum pH between 6.5 and 7.0 and was inhibited by cystatins from R. microplus. Moreover, rBbCp substrate specificity evaluation using a peptide phage display library showed a preference for Val > Leu > Phe. Finally, antibodies anti-rBbCp were able to interfere with B. bovis growth in vitro, which highlights the BbCp as a potential target for drug design. Instituto de Patobiología Fil: Lu, Stephen. Universidade Federal de São Paulo. Escola Paulista de Medicina. Department of Biochemistry; Brasil Fil: Ascencio, Mariano E. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina Fil: Torquato, Ricardo J.S. Universidade Federal de São Paulo. Escola Paulista de Medicina. Department of Biochemistry; Brasil Fil: Florin-Christensen, Monica. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiologia; Argentina Fil: Florin-Christensen, Monica. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Fil: Tanaka, Aparecida S. Universidade Federal de São Paulo. Escola Paulista de Medicina. Department of Biochemistry; Brasil Fil: Tanaka, Aparecida S. Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular; Brasil 2022-01-14T11:46:50Z 2022-01-14T11:46:50Z 2020-12 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/acceptedVersion http://hdl.handle.net/20.500.12123/11122 https://www.sciencedirect.com/science/article/abs/pii/S0300908420302182 0300-9084 https://doi.org/10.1016/j.biochi.2020.09.012 eng info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf Elsevier Biochimie 179 : 127-134 (December 2020) |
| spellingShingle | Cisteína Babesia bovis Peptidasas Proteínas Recombinantes Anticuerpos Cysteine Peptidases Recombinant Proteins Antibodies Lu, Stephen Ascencio, Mariano Torquato, Ricardo J.S. Florin-Christensen, Mónica Tanaka, Aparecida S. Kinetic characterization of a novel cysteine peptidase from the protozoan Babesia bovis, a potential target for drug design |
| title | Kinetic characterization of a novel cysteine peptidase from the protozoan Babesia bovis, a potential target for drug design |
| title_full | Kinetic characterization of a novel cysteine peptidase from the protozoan Babesia bovis, a potential target for drug design |
| title_fullStr | Kinetic characterization of a novel cysteine peptidase from the protozoan Babesia bovis, a potential target for drug design |
| title_full_unstemmed | Kinetic characterization of a novel cysteine peptidase from the protozoan Babesia bovis, a potential target for drug design |
| title_short | Kinetic characterization of a novel cysteine peptidase from the protozoan Babesia bovis, a potential target for drug design |
| title_sort | kinetic characterization of a novel cysteine peptidase from the protozoan babesia bovis a potential target for drug design |
| topic | Cisteína Babesia bovis Peptidasas Proteínas Recombinantes Anticuerpos Cysteine Peptidases Recombinant Proteins Antibodies |
| url | http://hdl.handle.net/20.500.12123/11122 https://www.sciencedirect.com/science/article/abs/pii/S0300908420302182 https://doi.org/10.1016/j.biochi.2020.09.012 |
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