RmKK, a tissue kallikrein inhibitor from Rhipicephalus microplus eggs

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dc.contributor.authorAbreu, Patricia A. [UNIFESP]
dc.contributor.authorSoares, Tatiane S. [UNIFESP]
dc.contributor.authorBuarque, Diego S. [UNIFESP]
dc.contributor.authorTorquato, Ricardo S. [UNIFESP]
dc.contributor.authorTanaka, Aparecida S. [UNIFESP]
dc.contributor.institutionUniversidade Federal de São Paulo (UNIFESP)
dc.date.accessioned2016-01-24T14:37:27Z
dc.date.available2016-01-24T14:37:27Z
dc.date.issued2014-06-20
dc.description.abstractRhipicephalus microplus is an important ectoparasite that is responsible for transmission of anaplasmosis and babesiosis to cattle. Tissue kallikrein inhibitors might play an important role in R. microplus eggs. in the present work, we purified and characterized, a tissue kallikrein inhibitor presents in R. microplus eggs (RmKK), a protein which contains two Kunitz domain in tandem. Purified inhibitor was confirmed by amino terminal determination and its dissociation constant (K-i) for bovine trypsin and porcine pancreatic kallikrein were 0.6 nM and 91.5 nM, respectively. Using a cDNA library from R. microplus midgut, we cloned the cDNA fragment encoding mature RmKK and expressed the protein in Pichia pastoris system. Recombinant RmKK was purified by ion exchange chromatography and presented molecular mass of 16.3 kDa by MALDI-TOF analysis. Moreover, RmKK showed a tight binding inhibition for serine proteases as bovine trypsin (K-i = 0.2 nM) and porcine pancreatic kallikrein (PPK) (K-i = 300 nM). We performed, for the first time, the characterization of a tissue kallikrein inhibitor presents in R. microplus eggs, which the transcript is produced in the adult female gut. BmKK seems to be the strongest PPK inhibitor among all BmTIs present in the eggs and larvae (Andreotti et al., 2001; Sasaki et al., 2004). This data suggests that BmKK may participate in the development of tick egg and larvae phase. (C) 2014 Elsevier Inc. All rights reserved.en
dc.description.affiliationUniversidade Federal de São Paulo, Dept Biochem, São Paulo, Brazil
dc.description.affiliationUnifespUniversidade Federal de São Paulo, Dept Biochem, São Paulo, 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.sponsorshipCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
dc.description.sponsorshipINCT - Entomologia Molecular
dc.description.sponsorshipIDFAPESP: 2005/03514-9
dc.description.sponsorshipIDFAPESP: 2012/03657-8
dc.format.extent69-73
dc.identifierhttp://dx.doi.org/10.1016/j.bbrc.2014.04.154
dc.identifier.citationBiochemical and Biophysical Research Communications. San Diego: Academic Press Inc Elsevier Science, v. 449, n. 1, p. 69-73, 2014.
dc.identifier.doi10.1016/j.bbrc.2014.04.154
dc.identifier.issn0006-291X
dc.identifier.urihttp://repositorio.unifesp.br/handle/11600/37879
dc.identifier.wosWOS:000337990700012
dc.language.isoeng
dc.publisherElsevier B.V.
dc.relation.ispartofBiochemical and Biophysical Research Communications
dc.rightsinfo:eu-repo/semantics/restrictedAccess
dc.rights.licensehttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dc.subjectRhipicephalus microplusen
dc.subjectEggsen
dc.subjectTissue kallikrein inhibitoren
dc.subjectSerine protease inhibitoren
dc.titleRmKK, a tissue kallikrein inhibitor from Rhipicephalus microplus eggsen
dc.typeinfo:eu-repo/semantics/article
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