Identification and molecular characterization of a novel Babesia orientalis thrombospondin-related anonymous protein (BoTRAP1)

Yu, Liang; Liu, Qin; Zhan, Xueyan; Huang, Yuan; Sun, Yali; Zheng, Na; Zhao, Yangnan; An, Xiaomeng; Li, Muxiao; Wang, Sen; Ao, Yangsiqi; Huang, Cuiqin; He, Li; Zhao, Junlong

doi:10.1186/s13071-018-3245-2

Cited by 9 publications

(13 citation statements)

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“…The involvement of members of the TRAP family during the erythrocyte invasion by merozoites has been previously demonstrated in B. bovis [6,27] and in other Babesia species [8][9][10]. Our results showing an important reduction in parasite invasion assays with anti-TRAP-1 antibodies support the hypothesis of the presence of surface-exposed neutralizing B-cell epitopes, which is in accordance with these previously published reports.…”

Section: Discussionsupporting

confidence: 93%

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Identification and characterization of a Babesia bigemina thrombospondin-related superfamily member, TRAP-1: a novel antigen containing neutralizing epitopes involved in merozoite invasion

et al. 2020

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Background Thrombospondin-related anonymous protein (TRAP) has been described as a potential vaccine candidate for several diseases caused by apicomplexan parasites. However, this protein and members of this family have not yet been characterized in Babesia bigemina, one of the most prevalent species causing bovine babesiosis. Methods The 3186-bp Babesia bigemina TRAP-1 (BbiTRAP-1) gene was identified by a bioinformatics search using the B. bovis TRAP-1 sequence. Members of the TRAP and TRAP-related protein families (TRP) were identified in Babesia and Theileria through a search of the TSP-1 adhesive domain, which is the hallmark motif in both proteins. Structural modeling and phylogenetic analysis were performed with the identified TRAP proteins. A truncated recombinant BbiTRAP-1 that migrates at approximately 107 kDa and specific antisera were produced and used in Western blot analysis and indirect fluorescent antibody tests (IFAT). B-cell epitopes with neutralizing activity in BbiTRAP-1 were defined by enzyme-linked immunosorbent assays (ELISA) and invasion assays. Results Three members of the TRAP family of proteins were identified in B. bigemina (BbiTRAP-1 to -3). All are type 1 transmembrane proteins containing the von Willebrand factor A (vWFA), thrombospondin type 1 (TSP-1), and cytoplasmic C-terminus domains, as well as transmembrane regions. The BbiTRAP-1 predicted structure also contains a metal ion-dependent adhesion site for interaction with the host cell. The TRP family in Babesia and Theileria species contains the canonical TSP-1 domain but lacks the vWFA domain and together with TRAP define a novel gene superfamily. A variable number of tandem repeat units are present in BbiTRAP-1 and could be used for strain genotyping. Western blot and IFAT analysis confirmed the expression of BbiTRAP-1 by blood-stage parasites. Partial recognition by a panel of sera from B. bigemina-infected cattle in ELISAs using truncated BbiTRAP-1 suggests that this protein is not an immunodominant antigen. Additionally, bovine anti-recombinant BbiTRAP-1 antibodies were found to be capable of neutralizing merozoite invasion in vitro. Conclusions We have identified the TRAP and TRP gene families in several Babesia and Theileria species and characterized BbiTRAP-1 as a novel antigen of B. bigemina. The functional relevance and presence of neutralization-sensitive B-cell epitopes suggest that BbiTRAP-1 could be included in tests for future vaccine candidates against B. bigemina.

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Section: Discussionsupporting

confidence: 93%

“…This protein was found to be directly involved in both the recognition and invasion processes of bovine erythrocytes. Other TRAPs have also been recently reported in Babesia orientalis and B. gibsoni [8][9][10].…”

Section: Introductionmentioning

confidence: 66%

Identification and characterization of a Babesia bigemina thrombospondin-related superfamily member, TRAP-1: a novel antigen containing neutralizing epitopes involved in merozoite invasion

et al. 2020

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“…The involvement of members of the TRAP family during the erythrocyte invasion by merozoites has been previously demonstrated in B. bovis (4,25) and in other Babesia species (6)(7)(8). Our results of an important reduction in parasite invasion assays with anti-TRAP-1 antibodies support the hypothesis of the presence of surface exposed neutralizing B-cell epitopes, which is in accordance to the abovementioned reports.…”

Section: Discussionsupporting

confidence: 92%

“…This protein was directly involved in both processes of recognition and invasion of bovine erythrocytes. Recently, other TRAP proteins have also been reported in B. orientalis and B. gibsoni (6)(7)(8).…”

mentioning

confidence: 98%

Identification and characterization of a Babesia bigemina thrombospondin-related superfamily member, TRAP-1: a novel antigen containing neutralizing epitopes involved in merozoite invasion

Montenegro

Paoletta

Ortiz

et al. 2020

Preprint

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Background The Thrombospondin-Related Anonymous Protein (TRAP) has been described as a potential vaccine candidate in several apicomplexan parasites. However, this protein and members of this family have not been characterized yet in Babesia bigemina, one of the most prevalent species causing bovine babesiosis.Methods The Babesia bigemina TRAP-1 (BbiTRAP-1) gene was identified by a bioinformatics search using the B. bovis TRAP-1 sequence. Members of the TRAP and TRAP-related protein family (TRP) families were identified in Babesia and Theileria through the search of the TSP-1 adhesive domain, the hallmark motif of TRAP proteins. Structural modelling and phylogenetic analysis were performed with the identified TRAP proteins. A truncated recombinant BbiTRAP-1 and specific antisera were produced, and used in Western blot analysis and indirect fluorescent antibody test (IFAT). B-cell epitopes with neutralizing activity in BbiTRAP-1 were defined by ELISA and invasion assays.Results TRAP family has 3 members in B. bigemina (BbiTRAP-1-3). All are type 1 transmembrane proteins containing the von Willebrand factor A (vWFA), thrombospondin type 1 (TSP-1) and cytoplasmic C-terminus domains along with transmembrane regions. The BbiTRAP-1 predicted structure also contains a metal ion-dependent adhesion site (MIDAS) for interaction with the host cell. The TRP family in Babesia and Theileria species contains the canonical TSP-1 domain but lack the vWFA domain,and together with TRAP define a novel gene superfamily. A variable number of tandem repeat units is present in BbiTRAP-1 and could be used for strain genotyping. Western blot and IFAT analysis confirmed expression of BbiTRAP-1 by blood stage parasites. Partial recognition by a panel of sera from B. bigemina infected cattle in ELISA using truncated BbiTRAP-1 suggests that this protein is not an immunodominant antigen. Additionally, bovine anti- recombinant BbiTRAP-1 antibodies were capable of neutralize merozoite invasion in vitro. Conclusions We identified the TRAP and TRP gene families in several Babesia and Theileria species, and characterized BbiTRAP-1 as a novel antigen of B. bigemina. The functional relevance and presence of neutralization-sensitive B-cell epitopes suggest that BbiTRAP-1 could be included in future vaccine candidates against B. bigemina.

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“…Like B. bovis , four TRAP genes also exist in B. orientalis and are considered as excellent vaccine candidates. BoTRAP1 has been identified which could be important for the interaction with ligands on the surface of the host cells and provide a theoretical basis for the discovery of novel Babesia vaccine candidate antigens [21].…”

Section: Introductionmentioning

confidence: 99%

Identification of a novel thrombospondin-related anonymous protein (BoTRAP2) from Babesia orientalis

Zhan

et al. 2019

Parasites Vectors

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Background The thrombospondin-related anonymous protein (TRAP) was first discovered in the sporozoite of Plasmodium falciparum and TRAP family proteins are secreted by micronemes and transported to the parasite surface to participate in the invasion process. Various TRAP proteins have been identified in apicomplexan protozoans, but there have been few reports about TRAP proteins in Babesia orientalis . Methods The functional domain of TRAP2 in B. orientalis was cloned, sequenced, characterized and compared to the TRAP sequences of related apicomplexan parasites. The functional domain of BoTRAP2 was truncated, named BoTRAP2-1, and then cloned into the pET-28a expression vector. Rabbit anti-rBoTRAP2-1 polyclonal antibody was produced by immunizing three rabbits. Western blot analysis was used to identify the native form and immunogenicity of BoTRAP2. The localization of BoTRAP2 was identified by indirect fluorescence assay (IFA). Results The amplified genes of BoTRAP2 are 2817 bp in length, encoding a functional domain of about 938 aa with two vWFA domains, one TSP domain and one transmembrane domain. The amino acid sequence of BoTRAP2 has a high similarity with that of B. bovis and B. gibsoni . The predicted tertiary structure of truncated BoTRAP2-1 confirmed that BoTRAP2 contains two vWFA domains and a TSP domain, the main functional areas of the protein. The native BoTRAP2 was identified from B. orientalis lysate by using rabbit polyclonal anti-rBoTRAP2-1. A band corresponding to rBoTRAP2-1 was detected by reaction with serum from a B. orientalis -infected water buffalo, indicating that the protein has a high immunogenicity. IFA showed that BoTRAP2 is mainly localized on the apical end of parasites by rabbit anti-rBoTRAP2-1 polyclonal serum. Conclusions The rBoTRAP2 could differentiate serum from B. orientalis -infected water buffalo and normal water buffalo, implicating that BoTRAP2 has high immunogenicity and could serve as a candidate antigen for diagnosis of B. orientalis infection in buffalo. Electronic supplementary material The online version of this article (10.1186/s13071-019-3457-0) contains supplementary material, which is available to authorized users.

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Identification and molecular characterization of a novel Babesia orientalis thrombospondin-related anonymous protein (BoTRAP1)

Cited by 9 publications

References 26 publications

Identification and characterization of a Babesia bigemina thrombospondin-related superfamily member, TRAP-1: a novel antigen containing neutralizing epitopes involved in merozoite invasion

Identification and characterization of a Babesia bigemina thrombospondin-related superfamily member, TRAP-1: a novel antigen containing neutralizing epitopes involved in merozoite invasion

Identification and characterization of a Babesia bigemina thrombospondin-related superfamily member, TRAP-1: a novel antigen containing neutralizing epitopes involved in merozoite invasion

Identification of a novel thrombospondin-related anonymous protein (BoTRAP2) from Babesia orientalis

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