Expression and functional characterization of boophilin, a thrombin inhibitor from Rhipicephalus (Boophilus) microplus midgut

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dc.contributor.authorSoares, Tatiane Sanches [UNIFESP]
dc.contributor.authorWatanabe, Renata Midori Okuta [UNIFESP]
dc.contributor.authorTanaka-Azevedo, Anita Mitico
dc.contributor.authorTorquato, Ricardo Jose Soares [UNIFESP]
dc.contributor.authorLu, Stephen [UNIFESP]
dc.contributor.authorFigueiredo, Ana Carvalho
dc.contributor.authorPereira, Pedro Jose Barbosa
dc.contributor.authorTanaka, Aparecida Sadae [UNIFESP]
dc.contributor.institutionUniversidade Federal de São Paulo (UNIFESP)
dc.contributor.institutionInst Butantan
dc.contributor.institutionUniv Porto
dc.date.accessioned2016-01-24T14:27:28Z
dc.date.available2016-01-24T14:27:28Z
dc.date.issued2012-07-06
dc.description.abstractRhipicephalus (Boophilus)microplus is an ectoparasite responsible for an important decrease in meat, milk and leather production, caused both by cattle blood loss and by the transmission of anaplasmosis and babesiosis. R. microplus is a rich source of serine protease inhibitors, including the trypsin inhibitors BmTI-A and BmTI-6, the subtilisin inhibitor BmSI, and the recently described thrombin inhibitor, boophilin. Boophilin is a double Kunitz-type thrombin inhibitor, with the unusual ability to form a ternary complex with a second (non-thrombin) serine proteinase molecule. the large-scale expression and purification of boophilin and of its isolated N-terminal (D1) domain in Pichia pastoris, its expression profile, and the effect of RNAi-mediated gene silencing in tick egg production are reported. Full-length boophilin and D1 were expressed at 21 and 37.5 mg/L of culture, respectively. Purified boophilin inhibited trypsin (K-i 0.65 nM), neutrophil elastase (K-i 21 nM) and bovine thrombin (K-i 57 pM), while D1 inhibited trypsin and neutrophil elastase (K-i of 2.0 and 129 nM, respectively), but not thrombin. Boophilin gene silencing using RNAi resulted in 20% reduction in egg weight production, suggesting that the expression of boophilin in this life stage would be important but not vital, probably due to functional overlap with other serine proteinase inhibitors in the midgut of R. microplus. Considering our data, Boophilin could be combining with other antigen in a vaccine production for tick control. (C) 2012 Elsevier B.V. All rights reserved.en
dc.description.affiliationUniversidade Federal de São Paulo, Escola Paulista Med, Dept Bioquim, BR-04044020 São Paulo, SP, Brazil
dc.description.affiliationInst Butantan, Lab Fisiopatol, São Paulo, SP, Brazil
dc.description.affiliationUniv Porto, IBMC, P-9150180 Oporto, Portugal
dc.description.affiliationUnifespUniversidade Federal de São Paulo, Escola Paulista Med, Dept Bioquim, BR-04044020 São Paulo, SP, Brazil
dc.description.sourceWeb of Science
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorshipConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.description.sponsorshipINCT-Entomologia Molecular
dc.description.sponsorshipFundacao para a Ciencia e a Tecnologia, Portugal
dc.description.sponsorshipEU-FEDER
dc.description.sponsorshipPOCI
dc.description.sponsorshipIDFAPESP: 05/03514-9
dc.description.sponsorshipIDFAPESP: 09/05405-3
dc.description.sponsorshipIDCNPq: 490574/2006-8
dc.description.sponsorshipIDFundacao para a Ciencia e a Tecnologia, Portugal: PTDC/BIA-PRO/70627/2006
dc.description.sponsorshipIDFundacao para a Ciencia e a Tecnologia, Portugal: REEQ/564/1310/2005
dc.description.sponsorshipID: SFR/BPD/46722/2008
dc.format.extent521-528
dc.identifierhttp://dx.doi.org/10.1016/j.vetpar.2012.01.027
dc.identifier.citationVeterinary Parasitology. Amsterdam: Elsevier B.V., v. 187, n. 3-4, p. 521-528, 2012.
dc.identifier.doi10.1016/j.vetpar.2012.01.027
dc.identifier.fileWOS000307323700024.pdf
dc.identifier.issn0304-4017
dc.identifier.urihttp://repositorio.unifesp.br/handle/11600/35089
dc.identifier.wosWOS:000307323700024
dc.language.isoeng
dc.publisherElsevier B.V.
dc.relation.ispartofVeterinary Parasitology
dc.rightsinfo:eu-repo/semantics/openAccess
dc.rights.licensehttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dc.subjectKunitz-type inhibitoren
dc.subjectTicken
dc.subjectAnticoagulanten
dc.subjectElastase inhibitoren
dc.subjectRNAi silencingen
dc.titleExpression and functional characterization of boophilin, a thrombin inhibitor from Rhipicephalus (Boophilus) microplus midguten
dc.typeinfo:eu-repo/semantics/article
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