Biochemical characterization of a Kunitz type inhibitor similar to dendrotoxins produced by Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) hemocytes

Show simple item record

dc.contributor.author Lima, Cassia A. [UNIFESP]
dc.contributor.author Torquato, Ricardo J. S. [UNIFESP]
dc.contributor.author Sasaki, Sergio D. [UNIFESP]
dc.contributor.author Justo, Giselle Z. [UNIFESP]
dc.contributor.author Tanaka, Aparecida S. [UNIFESP]
dc.date.accessioned 2016-01-24T13:59:19Z
dc.date.available 2016-01-24T13:59:19Z
dc.date.issued 2010-02-10
dc.identifier http://dx.doi.org/10.1016/j.vetpar.2009.09.030
dc.identifier.citation Veterinary Parasitology. Amsterdam: Elsevier B.V., v. 167, n. 2-4, p. 279-287, 2010.
dc.identifier.issn 0304-4017
dc.identifier.uri http://repositorio.unifesp.br/handle/11600/32267
dc.description.abstract A novel chymotrypsin inhibitor identified in fat body and hemocyte cDNA libraries of Boophilus microplus was named BmCI (B. microplus Chymotrypsin Inhibitor) (Genbank EU636772). the putative BmCI amino acid sequence presented a 22-residue-signal peptide and 58-residue-mature protein. BmCI amino acid sequence analysis allowed its classification as a Kunitz-BPTI inhibitor with six cysteine residues, a theoretical pI of 7.8, and the presence of Tyr at P1 position in the putative reactive site, suggesting inhibitory activity toward chymotrypsin. in this work, we reported the biochemical characterization of BmCI. the recombinant BmCI expressed in yeast Pichia pastoris was purified by size exclusion and reverse phase chromatographies. rBmCI expression yield was of I mg L-1 of culture. Purified rBmCI confirmed its chymotrypsin inhibitory activity with a low K-i (6.2 pM). the BmCI gene expression analysis by semi-quantitative RT-PCR indicated its transcription in the hemocytes, salivary gland and ovary. the cytotoxic activity of purified rBmCI was demonstrated in BALB/c 3T3 mouse fibroblasts. As assessed by the MTT reduction assay, rBMCI induced a dose-dependent decrease in 3T3 fibroblasts viability (IC50 = 8 mu M). Moreover, flow cytometry analysis revealed that rBmCI is able to induce apoptosis, whereas no effect was observed on cell cycle progression. in conclusion, we demonstrated that rBmCI is cytotoxic against mammalian cells and obtained evidence that this growth inhibition is caused by an apoptosis-inducing activity. (C) 2009 Published by Elsevier B.V. en
dc.description.sponsorship Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorship Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.description.sponsorship Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
dc.format.extent 279-287
dc.language.iso eng
dc.publisher Elsevier B.V.
dc.relation.ispartof Veterinary Parasitology
dc.rights Acesso restrito
dc.subject Kunitz-BPTI domain en
dc.subject Chymotrypsin inhibitor en
dc.subject Hemocytes en
dc.subject Ectoparasite en
dc.subject Hematophagous en
dc.subject Dendrotoxins en
dc.subject Cytotoxicity en
dc.subject Apoptosis en
dc.title Biochemical characterization of a Kunitz type inhibitor similar to dendrotoxins produced by Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) hemocytes en
dc.type Artigo
dc.rights.license http://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dc.contributor.institution Universidade Federal de São Paulo (UNIFESP)
dc.contributor.institution Universidade Estadual de Campinas (UNICAMP)
dc.description.affiliation Universidade Federal de São Paulo, Dept Bioquim, EPM, BR-04044020 São Paulo, Brazil
dc.description.affiliation Univ Estadual Campinas, Dept Bioquim, Inst Biol, BR-13083970 Campinas, SP, Brazil
dc.description.affiliationUnifesp Universidade Federal de São Paulo, Dept Bioquim, EPM, BR-04044020 São Paulo, Brazil
dc.description.sponsorshipID FAPESP: 05/03514-9
dc.description.sponsorshipID FAPESP: 06/52206-8
dc.identifier.doi 10.1016/j.vetpar.2009.09.030
dc.description.source Web of Science
dc.identifier.wos WOS:000274942600021



File

File Size Format View

There are no files associated with this item.

This item appears in the following Collection(s)

Show simple item record

Search


Browse

Statistics

My Account