Isolation and characterization of a new serine protease with thrombin-like activity (TLBm) from the venom of the snake Bothrops marajoensis

dc.creatorVilca-Quispe, A
dc.creatorPonce-Soto, LA
dc.creatorWinck, FV
dc.creatorMarangoni, S
dc.date2010
dc.dateAPR 1
dc.date2014-11-20T02:33:47Z
dc.date2015-11-26T16:03:32Z
dc.date2014-11-20T02:33:47Z
dc.date2015-11-26T16:03:32Z
dc.date.accessioned2018-03-28T22:52:46Z
dc.date.available2018-03-28T22:52:46Z
dc.identifierToxicon. Pergamon-elsevier Science Ltd, v. 55, n. 4, n. 745, n. 753, 2010.
dc.identifier0041-0101
dc.identifierWOS:000275705300009
dc.identifier10.1016/j.toxicon.2009.11.006
dc.identifierhttp://www.repositorio.unicamp.br/jspui/handle/REPOSIP/60800
dc.identifierhttp://www.repositorio.unicamp.br/handle/REPOSIP/60800
dc.identifierhttp://repositorio.unicamp.br/jspui/handle/REPOSIP/60800
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/1265288
dc.descriptionFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.descriptionThe thrombin-like serine protease TLBm from Bothrops marajoensis was isolated in one chromatographic step in reverse phase HPLC. Its molecular mass was 33239.95 Da, as based on the determined primary structure and confirmed experimentally by MALDI-TOF mass spectrometry (33332.5 Da) and it contains 12 half-cysteine residues. This TLBm exhibited high specificity for BA rho NA, Michaelis-Menten behavior with K(m) 2.3 x 10(-1) M and the V(max) 0.52 x 10(-1) nmoles rho-NA/lt/min for this substrate. TLBm also showed ability to coagulate bovine fibrinogen and was inhibited by soybean trypsin inhibitor, EDTA and S(Dm) from the serum of the species Didelphis marsupialis. The primary structure of TLBm showed the presence of His(45), Asp(103) and Ser(228) residues in the corresponding positions of the catalytic triad established in the serine proteases and Ser(228) are inhibited by phenylmethylsulfonyl fluoride (PMSF). Amino acid analysis showed a high content of Asp, Glu, Gly, Set, Ala and Pro as well as 12 half-cysteine residues and calculated pI of 6.47; TLBm presented 285 amino acid residues. In this work, we investigated the ability of TLBm to degrade fibrinogen and we observed that it is able to cause alpha- and beta-chain cleavage. Enzymatic as well as the platelet aggregation activities were strongly inhibited when incubated with PMSF, a specific inhibitor of serine protease. Also, TLBm induced platelet aggregation in washed and platelet-rich plasma, and in both cases, PMSF inhibited its activity. (C) 2009 Elsevier Ltd. All rights reserved.
dc.description55
dc.description4
dc.description745
dc.description753
dc.descriptionFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.descriptionMs Sc thesis of Augusto Vilca Quispe [06/54275-7]
dc.descriptionFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.descriptionMs Sc thesis of Augusto Vilca Quispe [06/54275-7]
dc.languageen
dc.publisherPergamon-elsevier Science Ltd
dc.publisherOxford
dc.publisherInglaterra
dc.relationToxicon
dc.relationToxicon
dc.rightsfechado
dc.rightshttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dc.sourceWeb of Science
dc.subjectBothrops marajoensis
dc.subjectSerine protease
dc.subjectThrombin-like enzyme
dc.subjectSnake venom
dc.subjectSerum from Didelphis marsupialis
dc.subjectAmino-acid-sequence
dc.subjectLachesis-muta-muta
dc.subjectExogenous Hemostatic Factors
dc.subjectMolecular-cloning
dc.subjectAtrox Venom
dc.subjectGland Transcriptome
dc.subjectActivating Enzyme
dc.subjectCoagulant Enzyme
dc.subjectPurification
dc.subjectViper
dc.titleIsolation and characterization of a new serine protease with thrombin-like activity (TLBm) from the venom of the snake Bothrops marajoensis
dc.typeArtículos de revistas


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