TY - JOUR
T1 - Unraveling the structure and function of CdcPDE: A novel phosphodiesterase from Crotalus durissus collilineatus snake venom
AU - de Oliveira, Isadora Sousa
AU - Pucca, Manuela Berto
AU - Wiezel, Gisele Adriano
AU - Cardoso, Iara Aimê
AU - de Castro Figueiredo Bordon, Karla
AU - Sartim, Marco Aurélio
AU - Kalogeropoulos, Konstantinos
AU - Ahmadi, Shirin
AU - Baiwir, Dominique
AU - Nonato, Maria Cristina
AU - Sampaio, Suely Vilela
AU - Laustsen, Andreas Hougaard
AU - auf dem Keller, Ulrich
AU - Quinton, Loïc
AU - Arantes, Eliane Candiani
PY - 2021
Y1 - 2021
N2 - 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.
AB - 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.
KW - Crotalus durissus collilineatus
KW - Phosphodiesterase
KW - Cytotoxicity
U2 - 10.1016/j.ijbiomac.2021.02.120
DO - 10.1016/j.ijbiomac.2021.02.120
M3 - Journal article
C2 - 33636276
SN - 0141-8130
VL - 178
SP - 180
EP - 192
JO - International Journal of Biological Macromolecules
JF - International Journal of Biological Macromolecules
ER -