Phosphoenolpyruvate carboxykinase from T. cruzi magnetic beads affinity-based screening assays on crude plant extracts from Brazilian Cerrado

Author(s)

do Amaral, Bruno S. ^{[1, 2]} ; da Silva, Larissa R. G. ^{[1, 3]} ; Valverde, Alessandra L. ^[3] ; de Sousa, Lorena R. F. ^[4] ; Severino, Richele P. ^[4] ; de Souza, Dulce H. F. ^[1] ; Cass, Quezia B. ^[1]

Affiliation

^[1] Univ Fed Sao Carlos, Dept Quim, Rodovia Washington Luis, Km 235, BR-13565905 Sao Carlos, SP - Brazil

^[2] Inst Fed Educ Ciencia & Tecnol Sao Paulo, Campus Pirituba, BR-05110000 Sao Paulo, SP - Brazil

^[3] Univ Fed Fluminense, Inst Quim, Programa Posgrad Quim, Niteroi, RJ - Brazil

^[4] Univ Fed Goias, Unidade Acad Especial Quim, BR-75704020 Catalao, Go - Brazil

• Document type: Journal article
• Source: Journal of Pharmaceutical and Biomedical Analysis; v. 193, JAN 30 2021.
• Web of Science Citations: 0

Abstract

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 K-Mapp value of 10 +/- 1 mu M 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. (C) 2020 Elsevier B.V. All rights reserved. (AU)

• FAPESP's process: 13/01710-1 - Enzyme ligand: new models of screening
• Grantee: Quezia Bezerra Cass
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 14/50249-8 - Green chemistry: sustainable synthetic methods employing benign solvents, safer reagents, and bio-renewable feedstock
• Grantee: Arlene Gonçalves Corrêa
• Support Opportunities: Research Grants - Research Centers in Engineering Program