Purification, Characterization and Evaluation of the Antitumoral Activity of a Phospholipase A2 from the Snake Bothrops moojeni .
Breno Emanuel Farias FrihlingAna Paula de Araújo BoletiCaio Fernando Ramalho de OliveiraSimone Camargo SanchesPedro Henrique de Oliveira CardosoNewton Valerio VerbisckMaria Lígia Rodrigues MacedoPaula Helena Santa RitaCristiano Marcelo Espinola CarvalhoLudovico MiglioloPublished in: Pharmaceuticals (Basel, Switzerland) (2022)
Nature presents a wide range of biomolecules with pharmacological potential, including venomous animal proteins. Among the protein components from snake venoms, phospholipases (PLA 2 ) are of great importance for the development of new anticancer compounds. Thus, we aimed to evaluate the PLA 2 anticancer properties from Bothrops moojeni venom. The crude venom was purified through three chromatographic steps, monitored by enzymatic activity and SDS-PAGE (12%). The purified PLA 2 denominated BmPLA2 had its molecular mass and N-terminal sequence identified by mass spectrometry and Edman degradation, respectively. BmPLA2 was assayed against human epithelial colorectal adenocarcinoma cells (Caco-2), human rhabdomyosarcoma cells (RD) and mucoepidermoid carcinoma of the lung (NCI-H292), using human fibroblast cells (MRC-5) and microglia cells (BV-2) as a cytotoxicity control. BmPLA2 presented 13,836 Da and a 24 amino acid-residue homologue with snake PLA 2 , which showed a 90% similarity with other Bothrops moojeni PLA 2 . BmPLA2 displayed an IC 50 of 0.6 µM against Caco-2, and demonstrated a selectivity index of 1.85 (compared to MRC-5) and 6.33 (compared to BV-2), supporting its selectivity for cancer cells. In conclusion, we describe a new acidic phospholipase, which showed antitumor activity and is a potential candidate in the development of new biotechnological tools.
Keyphrases
- induced apoptosis
- endothelial cells
- cell cycle arrest
- mass spectrometry
- amino acid
- endoplasmic reticulum stress
- squamous cell carcinoma
- oxidative stress
- induced pluripotent stem cells
- cell death
- high resolution
- cell proliferation
- nitric oxide
- risk assessment
- liquid chromatography
- small molecule
- single molecule
- climate change
- neuropathic pain
- hydrogen peroxide
- lipopolysaccharide induced
- ionic liquid
- capillary electrophoresis
- protein protein