| Author | Guedes, Herbert .L M. | |
| Author | Silva, Floriano P. | |
| Author | Netto, Carlos Correa | |
| Author | Salles, Cristiane M. C. de | |
| Author | Alexandre, Giani | |
| Author | Oliveira, Cristiano L. P. | |
| Author | Torriani, Iris | |
| Author | De Simone, Salvatore Giovanni | |
| Access date | 2019-03-14T12:03:48Z | |
| Available date | 2019-03-14T12:03:48Z | |
| Document date | 2008 | |
| Citation | GUEDES, Herbert L. M. et al. Structural characterization and low-resolution model of BJ-48, a thrombin-like enzyme from Bothrops jararacussu venom. Biophysical Chemistry, v.132, p.159–164, 2008. | pt_BR |
| ISSN | 0301-4622 | pt_BR |
| URI | https://www.arca.fiocruz.br/handle/icict/32069 | |
| Language | eng | pt_BR |
| Publisher | Elsevier | pt_BR |
| Rights | restricted access | |
| Subject in Portuguese | Veneno de cobra | pt_BR |
| Subject in Portuguese | Protease serina | pt_BR |
| Subject in Portuguese | Estrutura da solução | pt_BR |
| Subject in Portuguese | Forma de proteína | pt_BR |
| Subject in Portuguese | Mudanças conformacionais induzidas por pH | pt_BR |
| Title | Structural characterization and low-resolution model of BJ-48, a thrombin-like enzyme from Bothrops jararacussu venom | pt_BR |
| Type | Article | |
| DOI | 10.1016/j.bpc.2007.11.002 | |
| 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 A and a maximum dimension (D(max)) of 130 A as measured by small-angle X-ray scattering (SAXS) and a Stokes radius (SR) of 50 A 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 A and D(max)=82 A) and DLS (SR=30 A) 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 dichroism 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. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Bioquímica de Proteínas e Peptídeos. Rio de Janeiro, RJ. Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Bioquímica de Proteínas e Peptídeos. Rio de Janeiro, RJ. Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Bioquímica de Proteínas e Peptídeos. Rio de Janeiro, RJ. Brasil / Instituto Vital Brazil. Niterói, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Bioquímica de Proteínas e Peptídeos. Rio de Janeiro, RJ. Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Bioquímica de Proteínas e Peptídeos. Rio de Janeiro, RJ. Brasil. | pt_BR |
| Affilliation | Centro de Biologia Molecular Estrutural. Laboratório Nacional de Luz Síncrotron. Campinas, SP, Brasil / Universidade Estadual de Campinas. Instituto de Física. Campinas, SP, Brasil. | pt_BR |
| Affilliation | Centro de Biologia Molecular Estrutural. Laboratório Nacional de Luz Síncrotron. Campinas, SP, Brasil / Universidade Estadual de Campinas. Instituto de Física. Campinas, SP, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Bioquímica de Proteínas e Peptídeos. Rio de Janeiro, RJ. Brasil / Universidade Federal Fluminense. Instituto de Biologia. Departamento de Biologia Celular. Niterói, RJ, Brasil. | pt_BR |
| Subject | Snake venom | pt_BR |
| Subject | Serine protease | pt_BR |
| Subject | SVTLE | pt_BR |
| Subject | Solution structure | pt_BR |
| Subject | pH-induced conformational changes | pt_BR |
| Subject | Protein shape | pt_BR |
| Embargo date | 2022-01-01 | |
