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Molecular Characterization of a New Babesia bovis Thrombospondin-Related Anonymous Protein (BbTRAP2)

A gene encoding a Babesia bovis protein that shares significant degree of similarity to other apicomplexan thrombospondin-related anonymous proteins (TRAPs) was found in the genomic database and designated as BbTRAP2. Recombinant protein containing a conserved region of BbTRAP2 was produced in E. co...

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Autores principales: Terkawi, Mohamad Alaa, Ratthanophart, Jadsada, Salama, Akram, AbouLaila, Mahmoud, Asada, Masahito, Ueno, Akio, Alhasan, Hend, Guswanto, Azirwan, Masatani, Tatsunori, Yokoyama, Naoaki, Nishikawa, Yoshifumi, Xuan, Xuenan, Igarashi, Ikuo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3862764/
https://www.ncbi.nlm.nih.gov/pubmed/24349483
http://dx.doi.org/10.1371/journal.pone.0083305
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author Terkawi, Mohamad Alaa
Ratthanophart, Jadsada
Salama, Akram
AbouLaila, Mahmoud
Asada, Masahito
Ueno, Akio
Alhasan, Hend
Guswanto, Azirwan
Masatani, Tatsunori
Yokoyama, Naoaki
Nishikawa, Yoshifumi
Xuan, Xuenan
Igarashi, Ikuo
author_facet Terkawi, Mohamad Alaa
Ratthanophart, Jadsada
Salama, Akram
AbouLaila, Mahmoud
Asada, Masahito
Ueno, Akio
Alhasan, Hend
Guswanto, Azirwan
Masatani, Tatsunori
Yokoyama, Naoaki
Nishikawa, Yoshifumi
Xuan, Xuenan
Igarashi, Ikuo
author_sort Terkawi, Mohamad Alaa
collection PubMed
description A gene encoding a Babesia bovis protein that shares significant degree of similarity to other apicomplexan thrombospondin-related anonymous proteins (TRAPs) was found in the genomic database and designated as BbTRAP2. Recombinant protein containing a conserved region of BbTRAP2 was produced in E. coli. A high antigenicity of recombinant BbTRAP2 (rBbTRAP2) was observed with field B. bovis-infected bovine sera collected from geographically different regions of the world. Moreover, antiserum against rBbTRAP2 specifically reacted with the authentic protein by Western blot analysis and an indirect fluorescent antibody test. Three bands corresponding to 104-, 76-, and 44-kDa proteins were identified in the parasite lysates and two bands of 76- and 44-kDa proteins were detected in the supernatant of cultivated parasites, indicating that BbTRAP2 was proteolytically processed and shed into the culture. Apical and surface localizations of BbTRAP2 were observed in the intracellular and extracellular parasites, respectively, by confocal laser microscopic examination. Moreover, native BbTRAP2 was precipitated by bovine erythrocytes, suggesting its role in the attachment to erythrocytes. Furthermore, the specific antibody to rBbTRAP2 inhibited the growth of B. bovis in a concentration-dependent manner. Consistently, pre-incubation of the free merozoites with the antibody to rBbTRAP2 resulted in an inhibition of the parasite invasion into host erythrocytes. Interestingly, the antibody to rBbTRAP2 was the most inhibitive for the parasite’s growth as compared to those of a set of antisera produced against different recombinant proteins, including merozoite surface antigen 2c (BbMSA-2c), rhoptry-associated protein 1 C-terminal (BbRAP-1CT), and spherical body protein 1 (BbSBP-1). These results suggest that BbTRAP2 might be a potential candidate for development of a subunit vaccine against B. bovis infection.
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spelling pubmed-38627642013-12-17 Molecular Characterization of a New Babesia bovis Thrombospondin-Related Anonymous Protein (BbTRAP2) Terkawi, Mohamad Alaa Ratthanophart, Jadsada Salama, Akram AbouLaila, Mahmoud Asada, Masahito Ueno, Akio Alhasan, Hend Guswanto, Azirwan Masatani, Tatsunori Yokoyama, Naoaki Nishikawa, Yoshifumi Xuan, Xuenan Igarashi, Ikuo PLoS One Research Article A gene encoding a Babesia bovis protein that shares significant degree of similarity to other apicomplexan thrombospondin-related anonymous proteins (TRAPs) was found in the genomic database and designated as BbTRAP2. Recombinant protein containing a conserved region of BbTRAP2 was produced in E. coli. A high antigenicity of recombinant BbTRAP2 (rBbTRAP2) was observed with field B. bovis-infected bovine sera collected from geographically different regions of the world. Moreover, antiserum against rBbTRAP2 specifically reacted with the authentic protein by Western blot analysis and an indirect fluorescent antibody test. Three bands corresponding to 104-, 76-, and 44-kDa proteins were identified in the parasite lysates and two bands of 76- and 44-kDa proteins were detected in the supernatant of cultivated parasites, indicating that BbTRAP2 was proteolytically processed and shed into the culture. Apical and surface localizations of BbTRAP2 were observed in the intracellular and extracellular parasites, respectively, by confocal laser microscopic examination. Moreover, native BbTRAP2 was precipitated by bovine erythrocytes, suggesting its role in the attachment to erythrocytes. Furthermore, the specific antibody to rBbTRAP2 inhibited the growth of B. bovis in a concentration-dependent manner. Consistently, pre-incubation of the free merozoites with the antibody to rBbTRAP2 resulted in an inhibition of the parasite invasion into host erythrocytes. Interestingly, the antibody to rBbTRAP2 was the most inhibitive for the parasite’s growth as compared to those of a set of antisera produced against different recombinant proteins, including merozoite surface antigen 2c (BbMSA-2c), rhoptry-associated protein 1 C-terminal (BbRAP-1CT), and spherical body protein 1 (BbSBP-1). These results suggest that BbTRAP2 might be a potential candidate for development of a subunit vaccine against B. bovis infection. Public Library of Science 2013-12-13 /pmc/articles/PMC3862764/ /pubmed/24349483 http://dx.doi.org/10.1371/journal.pone.0083305 Text en © 2013 Terkawi et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Terkawi, Mohamad Alaa
Ratthanophart, Jadsada
Salama, Akram
AbouLaila, Mahmoud
Asada, Masahito
Ueno, Akio
Alhasan, Hend
Guswanto, Azirwan
Masatani, Tatsunori
Yokoyama, Naoaki
Nishikawa, Yoshifumi
Xuan, Xuenan
Igarashi, Ikuo
Molecular Characterization of a New Babesia bovis Thrombospondin-Related Anonymous Protein (BbTRAP2)
title Molecular Characterization of a New Babesia bovis Thrombospondin-Related Anonymous Protein (BbTRAP2)
title_full Molecular Characterization of a New Babesia bovis Thrombospondin-Related Anonymous Protein (BbTRAP2)
title_fullStr Molecular Characterization of a New Babesia bovis Thrombospondin-Related Anonymous Protein (BbTRAP2)
title_full_unstemmed Molecular Characterization of a New Babesia bovis Thrombospondin-Related Anonymous Protein (BbTRAP2)
title_short Molecular Characterization of a New Babesia bovis Thrombospondin-Related Anonymous Protein (BbTRAP2)
title_sort molecular characterization of a new babesia bovis thrombospondin-related anonymous protein (bbtrap2)
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3862764/
https://www.ncbi.nlm.nih.gov/pubmed/24349483
http://dx.doi.org/10.1371/journal.pone.0083305
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