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dc.contributor.authorLima, Cassia A.
dc.contributor.authorSasaki, Sergio D.
dc.contributor.authorTanaka, Aparecida S.
dc.date.accessioned2016-01-24T12:41:24Z
dc.date.available2016-01-24T12:41:24Z
dc.date.issued2006-08-18
dc.identifierhttp://dx.doi.org/10.1016/j.bbrc.2006.06.018
dc.identifier.citationBiochemical and Biophysical Research Communications. San Diego: Academic Press Inc Elsevier Science, v. 347, n. 1, p. 44-50, 2006.
dc.identifier.issn0006-291X
dc.identifier.urihttp://repositorio.unifesp.br/handle/11600/29090
dc.description.abstractThe bovine tick Rhipicephalus (Boophilus) microplus is a blood-sucking animal, which is responsible for Babesia spp and Anaplasma marginale transmission for cattle. From a B. microplus fat body cDNA library, 465 selected clones were sequenced randomly and resulted in 60 Contigs. An open reading frame (ORF) contains 98 amino acids named Bmcystatin, due to 70% amino acid identity to a classical type I cystatin from Ixodes scapularis tick (GenBank Accession No. DQ066227). the Bmcystatin amino acid sequence analysis showed two cysteine residues, theoretical pI of 5.92 and M-r of I I kDa. Bmcystatin gene was cloned in pET 26b vector and the protein expressed using bacteria Escherichia coli BL21 SI. Recombinant Bmcystatin (rBmcystatin) purified by affinity chromatography on Ni-NTA-aga-rose column and ionic exchange chromatography on HiTrap Q column presented molecular mass of I I kDa, by SDS-PAGE and the N-terminal amino acid sequenced revealed unprocessed N-terminal containing part of pelB signal sequence. Purified rBmcystatin showed to be a C I cysteine peptidase inhibitor with K-i value of 0.1 and 0.6 nM for human cathepsin L and VTDCE (vitellin degrading cysteine endopeptidase), respectively. the rBmcystatin expression analyzed by semi-quantitative RT-PCR confirmed the amplification of a specific DNA sequence (294 bp) in the fat body and ovary cDNA preparation. On the other hand, a protein band was detected in the fat body, ovary, and the salivary gland extracts using anti-Bmcystatin antibody by Western blot. the present results suggest a possible role of Bmcystatin in the ovary, even though the gene was cloned from the fat body, which could be another site of this protein synthesis. (c) 2006 Elsevier Inc. All rights reserved.en
dc.format.extent44-50
dc.language.isoeng
dc.publisherElsevier B.V.
dc.relation.ispartofBiochemical and Biophysical Research Communications
dc.rightsAcesso restrito
dc.subjectacarien
dc.subjectBoophilus microplusen
dc.subjectcysteine proteinase inhibitoren
dc.subjectcystatinen
dc.subjectstefin Ben
dc.titleBmeystatin, a cysteine proteinase inhibitor characterized from the tick Boophilus microplusen
dc.typeArtigo
dc.rights.licensehttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dc.contributor.institutionUniversidade Federal de São Paulo (UNIFESP)
dc.description.affiliationUniversidade Federal de São Paulo, Escola Paulista Med, Dept Biochem, BR-04044020 São Paulo, SP, Brazil
dc.description.affiliationUnifespUniversidade Federal de São Paulo, Escola Paulista Med, Dept Biochem, BR-04044020 São Paulo, SP, Brazil
dc.identifier.doi10.1016/j.bbrc.2006.06.018
dc.description.sourceWeb of Science
dc.identifier.wosWOS:000239198000007


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