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

Autor: Marco A. Sartim, Maria Cristina Nonato, Suely Vilela Sampaio, Gisele Adriano Wiezel, Shirin Ahmadi, Eliane Candiani Arantes, Konstantinos Kalogeropoulos, Karla de Castro Figueiredo Bordon, Andreas Hougaard Laustsen, Isadora Sousa de Oliveira, Dominique Baiwir, Loïc Quinton, Ulrich auf dem Keller, Manuela Berto Pucca, Iara Aimê Cardoso
Rok vydání: 2021
Předmět:
Zdroj: Repositório Institucional da USP (Biblioteca Digital da Produção Intelectual)
Universidade de São Paulo (USP)
instacron:USP
de Oliveira, I S, Pucca, M B, Wiezel, G A, Cardoso, I A, de Castro Figueiredo Bordon, K, Sartim, M A, Kalogeropoulos, K, Ahmadi, S, Baiwir, D, Nonato, M C, Sampaio, S V, Laustsen, A H, auf dem Keller, U, Quinton, L & Arantes, E C 2021, ' Unraveling the structure and function of Cdc PDE: A novel phosphodiesterase from Crotalus durissus collilineatus snake venom ', International Journal of Biological Macromolecules, vol. 178, pp. 180-192 . https://doi.org/10.1016/j.ijbiomac.2021.02.120
ISSN: 0141-8130
DOI: 10.1016/j.ijbiomac.2021.02.120
Popis: 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.
Databáze: OpenAIRE