Please use this identifier to cite or link to this item:
Type: Artigo de periódico
Title: Structural characterization and low-resolution model of BJ-48, a thrombin-like enzyme from Bothrops jararacussu venom
Author: Guedes, HLM
Silva, FP
Netto, CC
de Salles, CMC
Alexandre, G
Oliveria, CLP
Torriani, I
De Simone, SG
Abstract: Thrombin-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.
Subject: snake venom
serine protease
solution structure
pH-induced conformational changes
protein shape
Country: Holanda
Editor: Elsevier Science Bv
Citation: Biophysical Chemistry. Elsevier Science Bv, v. 132, n. 41700, n. 159, n. 164, 2008.
Rights: fechado
Identifier DOI: 10.1016/j.bpc.2007.11.002
Date Issue: 2008
Appears in Collections:Unicamp - Artigos e Outros Documentos

Files in This Item:
File Description SizeFormat 
WOS000252487800011.pdf523.17 kBAdobe PDFView/Open

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.