Characterization of a papain-like cysteine protease essential for the survival of Babesia ovis merozoites

Autores
Carletti, Tamara; Mesplet, Maria; Mira, Anabela; Weir, William; Shiels, Brian; Gonzalez Oliva, Abel; Schnittger, Leonhard; Florin-Christensen, Mónica; Barreto, Carmo
Año de publicación
2016
Idioma
inglés
Tipo de recurso
artículo
Estado
versión aceptada
Descripción
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
Fuente
Ticks and tick-borne diseases 7 (1) : 85-93. (February 2016)
Materia
Enfermedades de los Animales
Babesia Ovis
Papaína
Cisteína
Animal Diseases
Papain
Cysteine
Nivel de accesibilidad
acceso restringido
Condiciones de uso
Repositorio
INTA Digital (INTA)
Institución
Instituto Nacional de Tecnología Agropecuaria
OAI Identificador
oai:localhost:20.500.12123/1207
Acceder
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oai_identifier_str oai:localhost:20.500.12123/1207
network_acronym_str INTADig
repository_id_str l
network_name_str INTA Digital (INTA)
spelling Characterization of a papain-like cysteine protease essential for the survival of Babesia ovis merozoitesCarletti, TamaraMesplet, MariaMira, AnabelaWeir, WilliamShiels, BrianGonzalez Oliva, AbelSchnittger, LeonhardFlorin-Christensen, MónicaBarreto, CarmoEnfermedades de los AnimalesBabesia OvisPapaínaCisteínaAnimal DiseasesPapainCysteineBabesia 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 PatobiologíaFil: Carletti, Tamara. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; ArgentinaFil: 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; PortugalFil: Mesplet, Maria. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Cátedra de Enfermedades Infecciosas; ArgentinaFil: Mira, Anabela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Weir, William. Universityof Glasgow. College of Medical, Veterinary and Life Sciences; Reino UnidoFil: Shiels, Brian. Universityof Glasgow. College of Medical, Veterinary and Life Sciences; Reino UnidoFil: 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; PortugalFil: Schnittger, Leonhard. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiologia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Florin-Christensen, Monica. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiologia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina2017-09-13T14:48:02Z2017-09-13T14:48:02Z2016info:eu-repo/semantics/articleinfo:eu-repo/semantics/acceptedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/1207http://www.sciencedirect.com/science/article/pii/S1877959X1530008X1877-959Xhttps://doi.org/10.1016/j.ttbdis.2015.09.002Ticks and tick-borne diseases 7 (1) : 85-93. (February 2016)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repo/semantics/restrictedAccess2025-09-29T13:44:10Zoai:localhost:20.500.12123/1207instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-09-29 13:44:10.882INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv 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
spellingShingle Characterization of a papain-like cysteine protease essential for the survival of Babesia ovis merozoites
Carletti, Tamara
Enfermedades de los Animales
Babesia Ovis
Papaína
Cisteína
Animal Diseases
Papain
Cysteine
title_short 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_sort Characterization of a papain-like cysteine protease essential for the survival of Babesia ovis merozoites
dc.creator.none.fl_str_mv Carletti, Tamara
Mesplet, Maria
Mira, Anabela
Weir, William
Shiels, Brian
Gonzalez Oliva, Abel
Schnittger, Leonhard
Florin-Christensen, Mónica
Barreto, Carmo
author Carletti, Tamara
author_facet Carletti, Tamara
Mesplet, Maria
Mira, Anabela
Weir, William
Shiels, Brian
Gonzalez Oliva, Abel
Schnittger, Leonhard
Florin-Christensen, Mónica
Barreto, Carmo
author_role author
author2 Mesplet, Maria
Mira, Anabela
Weir, William
Shiels, Brian
Gonzalez Oliva, Abel
Schnittger, Leonhard
Florin-Christensen, Mónica
Barreto, Carmo
author2_role author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Enfermedades de los Animales
Babesia Ovis
Papaína
Cisteína
Animal Diseases
Papain
Cysteine
topic Enfermedades de los Animales
Babesia Ovis
Papaína
Cisteína
Animal Diseases
Papain
Cysteine
dc.description.none.fl_txt_mv 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
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.
publishDate 2016
dc.date.none.fl_str_mv 2016
2017-09-13T14:48:02Z
2017-09-13T14:48:02Z
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/acceptedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str acceptedVersion
dc.identifier.none.fl_str_mv 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
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
identifier_str_mv 1877-959X
dc.language.none.fl_str_mv eng
language eng
dc.rights.none.fl_str_mv info:eu-repo/semantics/restrictedAccess
eu_rights_str_mv restrictedAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv Ticks and tick-borne diseases 7 (1) : 85-93. (February 2016)
reponame:INTA Digital (INTA)
instname:Instituto Nacional de Tecnología Agropecuaria
reponame_str INTA Digital (INTA)
collection INTA Digital (INTA)
instname_str Instituto Nacional de Tecnología Agropecuaria
repository.name.fl_str_mv INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuaria
repository.mail.fl_str_mv tripaldi.nicolas@inta.gob.ar
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