Phosphoenolpyruvate carboxykinase from T. cruzi magnetic beads affinity-based screening assays on crude plant extracts from Brazilian Cerrado
Autor: | Alessandra L. Valverde, Richele P. Severino, Bruno S. do Amaral, Quezia B. Cass, Lorena R.F. de Sousa, Dulce Helena Ferreira de Souza, Larissa Ramos Guimarães da Silva |
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Rok vydání: | 2021 |
Předmět: |
Clinical Biochemistry
Pharmaceutical Science 01 natural sciences Malpighiaceae Analytical Chemistry Phosphoenolpyruvate chemistry.chemical_compound Liquid chromatography–mass spectrometry Vochysiaceae Drug Discovery Vanillic acid Humans Chagas Disease Spectroscopy biology Plant Extracts 010405 organic chemistry Magnetic Phenomena 010401 analytical chemistry Catechin biology.organism_classification 0104 chemical sciences Biochemistry chemistry Epimer Phosphoenolpyruvate carboxykinase Ebenaceae Brazil |
Zdroj: | Journal of Pharmaceutical and Biomedical Analysis. 193:113710 |
ISSN: | 0731-7085 |
Popis: | In T. cruzi, a causative agent of Chagas disease, phosphoenolpyruvate carboxykinase (TcPEPCK) is associated with carbohydrate catabolism. Due to its importance in the metabolism of the parasite, it has become a promising target for the development of new drugs against Chagas disease. Aiming to investigate different approaches for ligands screening, TcPEPCK was immobilized on amine-terminated magnetic beads (TcPEPCK-MB) and kinetically characterized by liquid chromatography tandem mass spectrometry activity assay with a KMapp value of 10 ± 1 μM to oxaloacetate as substrate. Natural products library affords highly diverse molecular frameworks through their secondary metabolites, herein a ligand fishing TcPEPCK-MB assay is described for prospecting ligands in four ethanolic extracts of Brazilian Cerrado plants: Qualea grandiflora (Vochysiaceae), Diospyros burchellii (Ebenaceae), Anadenanthera falcata (Fabaceae) and Byrsonima coccolobifolia (Malpighiaceae). The chemical characterization of eleven identified ligands was carried out by liquid chromatography tandem high-resolution mass spectrometry experiments. Senecic acid, syneilesinolide A, phytosphingosine and vanillic acid 4-glucopyranoside are herein reported for the first time for Q. grandiflora, D. burchellii, A. falcata, respectively. In addition, the specificity of the assay was observed since only catechin was fished out from the ethanolic extract of B. coccolobifolia leaves, despite the presence of epicatechin epimer. |
Databáze: | OpenAIRE |
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