Unraveling the structure and function of CdcPDE: A novel phosphodiesterase from Crotalus durissus collilineatus snake venom

Isadora Sousa de Oliveira, Manuela Berto Pucca, Gisele Adriano Wiezel, Iara Aimê Cardoso, Karla de Castro Figueiredo Bordon, Marco Aurélio Sartim, Konstantinos Kalogeropoulos, Shirin Ahmadi, Dominique Baiwir, Maria Cristina Nonato, Suely Vilela Sampaio, Andreas Hougaard Laustsen, Ulrich auf dem Keller, Loïc Quinton, Eliane Candiani Arantes*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

This study reports the isolation, structural, biochemical, and functional characterization of a novel phosphodiesterase from Crotalus durissus collilineatus venom (CdcPDE). CdcPDE was successfully isolated from whole venom using three chromatographic steps and represented 0.7% of total protein content. CdcPDE was inhibited by EDTA and reducing agents, demonstrating that metal ions and disulfide bonds are necessary for its enzymatic activity. The highest enzymatic activity was observed at pH 8-8.5 and 37 °C. Kinetic parameters indicated a higher affinity for the substrate bis(p-nitrophenyl) phosphate compared to others snake venom PDEs. Its structural characterization was done by the determination of the protein primary sequence by Edman degradation and mass spectrometry, and completed by the building of molecular and docking-based models. Functional in vitro assays showed that CdcPDE is capable of inhibiting platelet aggregation induced by adenosine diphosphate in a dose-dependent manner and demonstrated that CdcPDE is cytotoxic to human keratinocytes. CdcPDE was recognized by the crotalid antivenom produced by the Instituto Butantan. These findings demonstrate that the study of snake venom toxins can reveal new molecules that may be relevant in cases of snakebite envenoming, and that can be used as molecular tools to study pathophysiological processes due to their specific biological activities.
Original languageEnglish
JournalInternational Journal of Biological Macromolecules
Volume178
Pages (from-to)180-192
Number of pages13
ISSN0141-8130
DOIs
Publication statusPublished - 2021

Keywords

  • Crotalus durissus collilineatus
  • Phosphodiesterase
  • Cytotoxicity

Fingerprint Dive into the research topics of 'Unraveling the structure and function of <i>Cdc</i>PDE: A novel phosphodiesterase from <i>Crotalus durissus collilineatus</i> snake venom'. Together they form a unique fingerprint.

Cite this