Characterization of a papain-like cysteine protease essential for the survival of Babesia ovis merozoites
Babesia ovis, a tick-transmitted intraerythrocytic protozoan parasite, causes severe infections in small ruminants from Southern Europe, Middle East, and Northern Africa. With the aim of finding potential targets for the development of control methods against this parasite, sequence analysis of its...
| Autores principales: | , , , , , , , , |
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| Formato: | Artículo |
| Lenguaje: | Inglés |
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2017
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| Acceso en línea: | http://hdl.handle.net/20.500.12123/1207 http://www.sciencedirect.com/science/article/pii/S1877959X1530008X https://doi.org/10.1016/j.ttbdis.2015.09.002 |
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| author | Carletti, Tamara Mesplet, Maria Mira, Anabela Weir, William Shiels, Brian Gonzalez Oliva, Abel Schnittger, Leonhard Florin-Christensen, Mónica Barreto, Carmo |
| author_browse | Barreto, Carmo Carletti, Tamara Florin-Christensen, Mónica Gonzalez Oliva, Abel Mesplet, Maria Mira, Anabela Schnittger, Leonhard Shiels, Brian Weir, William |
| author_facet | Carletti, Tamara Mesplet, Maria Mira, Anabela Weir, William Shiels, Brian Gonzalez Oliva, Abel Schnittger, Leonhard Florin-Christensen, Mónica Barreto, Carmo |
| author_sort | Carletti, Tamara |
| collection | INTA Digital |
| description | Babesia ovis, a tick-transmitted intraerythrocytic protozoan parasite, causes severe infections in small ruminants from Southern Europe, Middle East, and Northern Africa. With the aim of finding potential targets for the development of control methods against this parasite, sequence analysis of its genome led to
the identification of four putative cysteine proteases of the C1A family. Orthology between B. ovis, B. bovis, T. annulata, and T. parva sequences showed that each B. ovis C1A peptidase sequence clustered within one of the four ortholog groups previously reported for these piroplasmids. The ortholog of bovipain-2 of B. bovis and falcipain-2 of Plasmodium falciparum, respectively, was designated “ovipain-2” and further characterized. In silico analysis showed that ovipain-2 has the typical topology of papain-like cysteine peptidases and a highly similar predicted three dimensional structure to bovipain-2 and falcipain-2, suggesting susceptibility to similar inhibitors. Immunoblotting using antibodies raised against a recombinant form of ovipain-2 (r-ovipain-2) demonstrated expression of ovipain-2 in in vitro cultured B. ovis merozoites.
By immunofluorescence, these antibodies reacted with merozoites and stained the cytoplasm of infected erythrocytes. This suggests that ovipain-2 is secreted by the parasite and could be involved in intra- and extracellular digestion of hemoglobin and/or cleavage of erythrocyte proteins facilitating parasite egress. A significant reduction in the percentage of parasitized erythrocytes was obtained upon incubation of B. ovis in vitro cultures with anti-r-ovipain-2 antibodies, indicating an important functional role for ovipain-2 in the intra erythrocytic development cycle of this parasite. Finally, studies of the reactivity of sera from B. ovis-positive and negative sheep against r-ovipain-2 showed that this protease is expressed in vivo, and can be recognized by host antibodies. The results of this study suggest that ovipain-2 constitutes a potential target for immunotherapies and drug development against ovine babesiosis. |
| format | Artículo |
| id | INTA1207 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| record_format | dspace |
| spelling | INTA12072019-01-24T17:44:17Z Characterization of a papain-like cysteine protease essential for the survival of Babesia ovis merozoites Carletti, Tamara Mesplet, Maria Mira, Anabela Weir, William Shiels, Brian Gonzalez Oliva, Abel Schnittger, Leonhard Florin-Christensen, Mónica Barreto, Carmo Enfermedades de los Animales Babesia Ovis Papaína Cisteína Animal Diseases Papain Cysteine Babesia ovis, a tick-transmitted intraerythrocytic protozoan parasite, causes severe infections in small ruminants from Southern Europe, Middle East, and Northern Africa. With the aim of finding potential targets for the development of control methods against this parasite, sequence analysis of its genome led to the identification of four putative cysteine proteases of the C1A family. Orthology between B. ovis, B. bovis, T. annulata, and T. parva sequences showed that each B. ovis C1A peptidase sequence clustered within one of the four ortholog groups previously reported for these piroplasmids. The ortholog of bovipain-2 of B. bovis and falcipain-2 of Plasmodium falciparum, respectively, was designated “ovipain-2” and further characterized. In silico analysis showed that ovipain-2 has the typical topology of papain-like cysteine peptidases and a highly similar predicted three dimensional structure to bovipain-2 and falcipain-2, suggesting susceptibility to similar inhibitors. Immunoblotting using antibodies raised against a recombinant form of ovipain-2 (r-ovipain-2) demonstrated expression of ovipain-2 in in vitro cultured B. ovis merozoites. By immunofluorescence, these antibodies reacted with merozoites and stained the cytoplasm of infected erythrocytes. This suggests that ovipain-2 is secreted by the parasite and could be involved in intra- and extracellular digestion of hemoglobin and/or cleavage of erythrocyte proteins facilitating parasite egress. A significant reduction in the percentage of parasitized erythrocytes was obtained upon incubation of B. ovis in vitro cultures with anti-r-ovipain-2 antibodies, indicating an important functional role for ovipain-2 in the intra erythrocytic development cycle of this parasite. Finally, studies of the reactivity of sera from B. ovis-positive and negative sheep against r-ovipain-2 showed that this protease is expressed in vivo, and can be recognized by host antibodies. The results of this study suggest that ovipain-2 constitutes a potential target for immunotherapies and drug development against ovine babesiosis. Inst. de Patobiología Fil: Carletti, Tamara. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina Fil: Barreto, Carmo. Universidade Nova de Lisboa. Instituto de Tecnologia Química e Biológica. Laboratório de Diagnóstico Biomolecular; Portugal. Universidade Nova de Lisboa. Instituto de Biologia Experimental e Tecnologica; Portugal Fil: Mesplet, Maria. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Cátedra de Enfermedades Infecciosas; Argentina Fil: Mira, Anabela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Weir, William. Universityof Glasgow. College of Medical, Veterinary and Life Sciences; Reino Unido Fil: Shiels, Brian. Universityof Glasgow. College of Medical, Veterinary and Life Sciences; Reino Unido Fil: Gonzalez Oliva, Abel. Universidade Nova de Lisboa. Instituto de Tecnologia Química e Biológica. Laboratório de Diagnóstico Biomolecular; Portugal. Universidade Nova de Lisboa. Instituto de Biologia Experimental e Tecnologica; Portugal Fil: Schnittger, Leonhard. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiologia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Florin-Christensen, Monica. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiologia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina 2017-09-13T14:48:02Z 2017-09-13T14:48:02Z 2016 info:eu-repo/semantics/article info:eu-repo/semantics/acceptedVersion info:ar-repo/semantics/artículo http://hdl.handle.net/20.500.12123/1207 http://www.sciencedirect.com/science/article/pii/S1877959X1530008X 1877-959X https://doi.org/10.1016/j.ttbdis.2015.09.002 eng info:eu-repo/semantics/restrictedAccess application/pdf Ticks and tick-borne diseases 7 (1) : 85-93. (February 2016) |
| spellingShingle | Enfermedades de los Animales Babesia Ovis Papaína Cisteína Animal Diseases Papain Cysteine Carletti, Tamara Mesplet, Maria Mira, Anabela Weir, William Shiels, Brian Gonzalez Oliva, Abel Schnittger, Leonhard Florin-Christensen, Mónica Barreto, Carmo Characterization of a papain-like cysteine protease essential for the survival of Babesia ovis merozoites |
| title | Characterization of a papain-like cysteine protease essential for the survival of Babesia ovis merozoites |
| title_full | Characterization of a papain-like cysteine protease essential for the survival of Babesia ovis merozoites |
| title_fullStr | Characterization of a papain-like cysteine protease essential for the survival of Babesia ovis merozoites |
| title_full_unstemmed | Characterization of a papain-like cysteine protease essential for the survival of Babesia ovis merozoites |
| title_short | Characterization of a papain-like cysteine protease essential for the survival of Babesia ovis merozoites |
| title_sort | characterization of a papain like cysteine protease essential for the survival of babesia ovis merozoites |
| topic | Enfermedades de los Animales Babesia Ovis Papaína Cisteína Animal Diseases Papain Cysteine |
| url | http://hdl.handle.net/20.500.12123/1207 http://www.sciencedirect.com/science/article/pii/S1877959X1530008X https://doi.org/10.1016/j.ttbdis.2015.09.002 |
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