[en] Vascular endothelial growth factors (VEGFs) are crucial molecules involved in the modulation of angiogenesis. Snake venom-derived VEGFs (svVEGFs) are known to contribute significantly to the envenoming due to their capacity of increasing vascular permeability. In our work, we isolated and analyzed the biochemical and functional properties of the VEGF from Crotalus durissus collilineatus venom (CdcVEGF). The venom was fractionated by reversed phase chromatography on FPLC system (Fast Protein Liquid Chromatography) and the eluted fractions were submitted to an ELISA assay using an anti-VEGF-F antibody, for identification of svVEGF. Positive fractions for svVEGF were submitted to SDS-PAGE and to an anion exchange chromatography to isolate the molecule. The subfractions were analyzed by ELISA and SDS-PAGE and six of them presented svVEGFs, named CdcVEGF1 (Q23-3), CdcVEGF2 (Q24-3), CdcVEGF3 (Q24-4), CdcVEGF4 (Q25-3), CdcVEGF5 (Q25-4), and CdcVEGF6 (Q25-5). Their structural characterization was accomplished by mass spectrometry analysis using MALDI-TOF to determine their molecular masses and UPLC-ESI-QTOF to determine their amino acid sequence. Interestingly, all isolated CdcVEGFs induced angiogenesis on HUVEC cells through tube formation on Matrigel when compared to culture medium (negative control). Moreover, CdcVEGF2 and CdcVEGF3 also induced a significant increase in tube formation when compared to the positive control (basic fibroblast growth factor - bFGF). Additionally, crotalid antivenom produced by the Instituto Butantan was able to recognize CdcVEGFs, demonstrating to be immunogenic. This study demonstrates that snake venom cocktail can reveal novel and important molecules, which are potential molecular tools to study diverse biological processes, such as angiogenesis.
Ferreira, Isabela Gobbo; Department of BioMolecular Sciences, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil
Pucca, Manuela Berto; Medical School, Federal University of Roraima, Boa Vista, RR, Brazil
Cardoso, Iara Aimê; Department of BioMolecular Sciences, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil
de Castro Figueiredo Bordon, Karla; Department of BioMolecular Sciences, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil
Wiezel, Gisele Adriano; Department of BioMolecular Sciences, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil
Gobbi Amorim, Fernanda ; Université de Liège - ULiège > Département de chimie (sciences) > Laboratoire de spectrométrie de masse (L.S.M.)
Rodrigues, Renata Santos; Institute of Biotechnology, Federal University of Uberlandia, UFU, Uberlandia, MG, Brazil
de Melo Rodrigues, Veridiana; Institute of Biotechnology, Federal University of Uberlandia, UFU, Uberlandia, MG, Brazil
Lucia de Campos Brites, Vera; Institute of Biology, Sector of Reptiles, Federal University of Uberlandia, UFU, Uberlandia, MG, Brazil
Rosa, José César; Protein Chemistry Center and Department of Molecular and Cell Biology and Pathogenic Bioagents, School of Medicine of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil
Lopes, Daiana Silva; Institute Multidisciplinary in Health, Universidade Federal da Bahia, Vitoria da Conquista, BA, Brazil
Arantes, Eliane Candiani; Department of BioMolecular Sciences, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil. Electronic address: ecabraga@fcfrp.usp.br
Language :
English
Title :
Insights into structure and function of CdcVEGFs, the vascular endothelial growth factor from Crotalus durissus collilineatus snake venom.
This work was funded by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior ( CAPES , Coordination for the Improvement of Higher Education Personnel , scholarship to IGF, finance code 001), Conselho Nacional de Desenvolvimento Científico e Tecnológico ( CNPq , The National Council for Scientific and Technological Development, scholarship to MBP 307184/2020-0 and grant 306479/2017-6), Fundação de Amparo à Pesquisa do Estado de São Paulo ( FAPESP , São Paulo Research Foundation, grants 2011/23236-4, 2019/10173-6 and scholarship to GAW 2017/00586-6, and the Nucleus for Research on Animal Toxins ( NAP-TOXAN-USP , grant 12–125432.1.3).
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