Structural characterization and low-resolution model of BJ-48, a thrombin-like enzyme from Bothrops jararacussu venom

dc.creatorGuedes, HLM
dc.creatorSilva, FP
dc.creatorNetto, CC
dc.creatorde Salles, CMC
dc.creatorAlexandre, G
dc.creatorOliveria, CLP
dc.creatorTorriani, I
dc.creatorDe Simone, SG
dc.date2008
dc.dateFEB
dc.date2014-11-15T19:24:42Z
dc.date2015-11-26T17:21:20Z
dc.date2014-11-15T19:24:42Z
dc.date2015-11-26T17:21:20Z
dc.date.accessioned2018-03-29T00:08:50Z
dc.date.available2018-03-29T00:08:50Z
dc.identifierBiophysical Chemistry. Elsevier Science Bv, v. 132, n. 41700, n. 159, n. 164, 2008.
dc.identifier0301-4622
dc.identifierWOS:000252487800011
dc.identifier10.1016/j.bpc.2007.11.002
dc.identifierhttp://www.repositorio.unicamp.br/jspui/handle/REPOSIP/79176
dc.identifierhttp://www.repositorio.unicamp.br/handle/REPOSIP/79176
dc.identifierhttp://repositorio.unicamp.br/jspui/handle/REPOSIP/79176
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/1283341
dc.descriptionThrombin-like enzymes (TLEs) are important components of snake venoms due to their involvement in coagulopathies occurring on envenoming. Structural characterization of this group of serine proteases is of utmost importance for better understanding their unique properties. However, the high carbohydrate content of some members of this group prevents successful crystallization for structural determination. Circumventing this difficulty, the structure of BJ-48, a highly glycosylated TLE from Bothrops jararacussu venom, was studied in solution. At pH 8.0, where the enzyme displays maximum activity, BJ-48 has a radius of gyration (Rg) of 37 angstrom and a maximum dimension (D-max) of 130 angstrom as measured by small-angle X-ray scattering (SAXS) and a Stokes radius (SR) of 50 angstrom according to dynamic light scattering (DLS) data. At the naturally more acidic pH (6.0) of the B. jararacussu venom BJ-48 behaves as a more compact particle as evidenced by SAXS (R-g= 27.9 angstrom and D-max = 82 angstrom) and DLS (SR=30 angstrom) data. In addition, Kratky plot analysis indicates a rigid shape at pH 8.0 and a flexible shape at pH 6.0. On the other hand, the center of mass of intrinsic fluorescence was not changed while varying pH, possibly indicating the absence of fluorescent amino acids in the regions affected by pH variation. Circular dichroisin experiments carried out with BJ-48 indicate a substantially random coiled secondary structure that is not affected by pH. Low-resolution model of BJ-48 presented a prolate elongated shape at pH 8.0 and a U-shape at 6.0. BJ-48 tertiary structure at pH 6.0 was maintained on heating up to 52 degrees C and was completely lost at 75 degrees C. The possible existence of two pH-induced folding states for BJ-48 and its importance for the biological role and stability of this enzyme was discussed. (C) 2007 Elsevier B.V. All rights reserved.
dc.description132
dc.description41700
dc.description159
dc.description164
dc.languageen
dc.publisherElsevier Science Bv
dc.publisherAmsterdam
dc.publisherHolanda
dc.relationBiophysical Chemistry
dc.relationBiophys. Chem.
dc.rightsfechado
dc.rightshttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dc.sourceWeb of Science
dc.subjectsnake venom
dc.subjectserine protease
dc.subjectSVTLE
dc.subjectsolution structure
dc.subjectpH-induced conformational changes
dc.subjectprotein shape
dc.subjectSmall-angle Scattering
dc.subjectHuman Alpha-thrombin
dc.subjectSerine Proteinases
dc.subjectCrystal-structure
dc.subjectSnake-venoms
dc.subjectActive-site
dc.subjectTrypsin
dc.subjectProteases
dc.subjectBovine
dc.subjectArg
dc.titleStructural characterization and low-resolution model of BJ-48, a thrombin-like enzyme from Bothrops jararacussu venom
dc.typeArtículos de revistas


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