| Full text | |
| Author(s): |
Sartim, Marco A.
;
Pinheiro, Matheus P.
;
de Padua, Ricardo A. P.
;
Sampaio, Suely V.
;
Cristina Nonato, M.
Total Authors: 5
|
| Document type: | Journal article |
| Source: | Toxicon; v. 126, p. 59-69, FEB 2017. |
| Web of Science Citations: | 6 |
| Abstract | |
BJcuL is a snake venom galactoside-binding lectin (SVgalL) isolated from Bothrops jararacussu and is involved in a wide variety of biological activities including triggering of pro-inflammatory response, disruption of microbial biofilm structure and induction of apoptosis. In the present work, we determined the crystallographic structure of BJcuL, the first holo structure of a SVgalL, and introduced the fluorescence-based thermal stability assay (Thermofluor) as a tool for screening and characterization of the binding mechanism of SVgalL ligands. BJcuL structure revealed the existence of a porous and flexible decameric arrangement composed of disulfide-linked dimers related by a five-fold symmetry. Each monomer contains the canonical carbohydrate recognition domain, a calcium ion required for BJcuL lectinic activity and a sodium ion required for protein stabilization. BJcuL thermostability was found to be induced by calcium ion and galactoside sugars which exhibit hyperbolic saturation profiles dependent on ligand concentration. Serendipitously, the gentamicin group of aminoglycoside antibiotics (gAGAs) was also identified as BJcuL ligands. On contrast, gAGAs exhibited a sigmoidal saturation profile compatible with a cooperative mechanism of binding. Thermofluor, hemagglutination inhibition assay and molecular docking strategies were used to identify a distinct binding site in BJcuL localized at the dimeric interface near the fully conserved intermolecular Cys86-Cys86 disulfide bond. The hybrid approach used in the present work provided novel insights into structural behavior and functional diversification of SVgaLs. (C) 2016 Elsevier Ltd. All rights reserved. (AU) | |
| FAPESP's process: | 11/23236-4 - Native and recombinant animal toxins: functional, structural and molecular analysis |
| Grantee: | Suely Vilela |
| Support Opportunities: | Research Projects - Thematic Grants |
| FAPESP's process: | 14/19127-3 - Effect of Calloselasma rhodostoma and Bothrops moojeni LAAO on apoptotic machinery of JAK2V617F positive strains of myeloproliferative neoplasms |
| Grantee: | Thaís Fontanezi Maciel |
| Support Opportunities: | Scholarships in Brazil - Scientific Initiation |
| FAPESP's process: | 14/11266-4 - Structural and biophysical characterization of animal toxins |
| Grantee: | Ricardo Augusto Pereira de Pádua |
| Support Opportunities: | Scholarships in Brazil - Post-Doctoral |