Bmeystatin, a cysteine proteinase inhibitor characterized from the tick Boophilus microplus

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dc.contributor.author Lima, Cassia A.
dc.contributor.author Sasaki, Sergio D.
dc.contributor.author Tanaka, Aparecida S.
dc.date.accessioned 2016-01-24T12:41:24Z
dc.date.available 2016-01-24T12:41:24Z
dc.date.issued 2006-08-18
dc.identifier http://dx.doi.org/10.1016/j.bbrc.2006.06.018
dc.identifier.citation Biochemical and Biophysical Research Communications. San Diego: Academic Press Inc Elsevier Science, v. 347, n. 1, p. 44-50, 2006.
dc.identifier.issn 0006-291X
dc.identifier.uri http://repositorio.unifesp.br/handle/11600/29090
dc.description.abstract The 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.extent 44-50
dc.language.iso eng
dc.publisher Elsevier B.V.
dc.relation.ispartof Biochemical and Biophysical Research Communications
dc.rights Acesso restrito
dc.subject acari en
dc.subject Boophilus microplus en
dc.subject cysteine proteinase inhibitor en
dc.subject cystatin en
dc.subject stefin B en
dc.title Bmeystatin, a cysteine proteinase inhibitor characterized from the tick Boophilus microplus 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.description.affiliation Universidade Federal de São Paulo, Escola Paulista Med, Dept Biochem, BR-04044020 São Paulo, SP, Brazil
dc.description.affiliationUnifesp Universidade Federal de São Paulo, Escola Paulista Med, Dept Biochem, BR-04044020 São Paulo, SP, Brazil
dc.identifier.doi 10.1016/j.bbrc.2006.06.018
dc.description.source Web of Science
dc.identifier.wos WOS:000239198000007



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