Kinetic characterization of a novel cysteine peptidase from the protozoan Babesia bovis, a potential target for drug design

Autores
Lu, Stephen; Ascencio, Mariano; Torquato, Ricardo J.S.; Florin-Christensen, Mónica; Tanaka, Aparecida S.
Año de publicación
2020
Idioma
inglés
Tipo de recurso
artículo
Estado
versión aceptada
Descripción
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
Fuente
Biochimie 179 : 127-134 (December 2020)
Materia
Cisteína
Babesia bovis
Peptidasas
Proteínas Recombinantes
Anticuerpos
Cysteine
Peptidases
Recombinant Proteins
Antibodies
Nivel de accesibilidad
acceso abierto
Condiciones de uso
http://creativecommons.org/licenses/by-nc-sa/4.0/
Repositorio
INTA Digital (INTA)
Institución
Instituto Nacional de Tecnología Agropecuaria
OAI Identificador
oai:localhost:20.500.12123/11122
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network_name_str INTA Digital (INTA)
spelling Kinetic characterization of a novel cysteine peptidase from the protozoan Babesia bovis, a potential target for drug designLu, StephenAscencio, MarianoTorquato, Ricardo J.S.Florin-Christensen, MónicaTanaka, Aparecida S.CisteínaBabesia bovisPeptidasasProteínas RecombinantesAnticuerposCysteinePeptidasesRecombinant ProteinsAntibodiesC1A 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 PatobiologíaFil: Lu, Stephen. Universidade Federal de São Paulo. Escola Paulista de Medicina. Department of Biochemistry; BrasilFil: Ascencio, Mariano E. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; ArgentinaFil: Torquato, Ricardo J.S. Universidade Federal de São Paulo. Escola Paulista de Medicina. Department of Biochemistry; BrasilFil: Florin-Christensen, Monica. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiologia; ArgentinaFil: 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; BrasilFil: Tanaka, Aparecida S. Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular; BrasilElsevier2022-01-14T11:46:50Z2022-01-14T11:46:50Z2020-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/acceptedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/11122https://www.sciencedirect.com/science/article/abs/pii/S03009084203021820300-9084https://doi.org/10.1016/j.biochi.2020.09.012Biochimie 179 : 127-134 (December 2020)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)2025-09-29T13:45:27Zoai:localhost:20.500.12123/11122instacron: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 13:45:27.81INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv 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
spellingShingle Kinetic characterization of a novel cysteine peptidase from the protozoan Babesia bovis, a potential target for drug design
Lu, Stephen
Cisteína
Babesia bovis
Peptidasas
Proteínas Recombinantes
Anticuerpos
Cysteine
Peptidases
Recombinant Proteins
Antibodies
title_short 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_sort Kinetic characterization of a novel cysteine peptidase from the protozoan Babesia bovis, a potential target for drug design
dc.creator.none.fl_str_mv Lu, Stephen
Ascencio, Mariano
Torquato, Ricardo J.S.
Florin-Christensen, Mónica
Tanaka, Aparecida S.
author Lu, Stephen
author_facet Lu, Stephen
Ascencio, Mariano
Torquato, Ricardo J.S.
Florin-Christensen, Mónica
Tanaka, Aparecida S.
author_role author
author2 Ascencio, Mariano
Torquato, Ricardo J.S.
Florin-Christensen, Mónica
Tanaka, Aparecida S.
author2_role author
author
author
author
dc.subject.none.fl_str_mv Cisteína
Babesia bovis
Peptidasas
Proteínas Recombinantes
Anticuerpos
Cysteine
Peptidases
Recombinant Proteins
Antibodies
topic Cisteína
Babesia bovis
Peptidasas
Proteínas Recombinantes
Anticuerpos
Cysteine
Peptidases
Recombinant Proteins
Antibodies
dc.description.none.fl_txt_mv 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
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.
publishDate 2020
dc.date.none.fl_str_mv 2020-12
2022-01-14T11:46:50Z
2022-01-14T11:46:50Z
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/20.500.12123/11122
https://www.sciencedirect.com/science/article/abs/pii/S0300908420302182
0300-9084
https://doi.org/10.1016/j.biochi.2020.09.012
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
identifier_str_mv 0300-9084
dc.language.none.fl_str_mv eng
language eng
dc.rights.none.fl_str_mv 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)
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv Biochimie 179 : 127-134 (December 2020)
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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score 12.559606

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