Departament de Química Analítica, Facultat de Química, Universitat de València, Dr. Moliner 50, 46100 Burjassot, Valencia, Spain.
Anal Chem. 2013 Apr 16;85(8):4014-21. doi: 10.1021/ac303746k. Epub 2013 Apr 2.
The interaction between heme and ligands is the basis for a variety of tests aimed at the discovery of antiplasmodial molecules. Two electrochemical methods for the screening of molecules with potential antimalarial activity through heme-binding mechanism are described. The first method is applicable to lipophilic environment, by using solution phase electrochemistry in DMSO solutions of Fe(III)-heme plus the tested compounds at carbon electrodes. This method provides well-defined voltammetric signals, characteristic of the heme-ligand (L) interaction. The second method involves aqueous media at biological pH and the use of voltammetry of immobilized particles, by means of microparticulate films of the tested compounds immersed into Fe(III)-heme solutions with no need of prior incubation. These methodologies are applied to the testing of heme-binding activity in macromolecular level systems like hemoglobin, or much more complex mixtures like total blood, erythrocytes, or hemolyzed samples.
血红素与配体的相互作用是发现抗疟分子的各种测试的基础。本文描述了两种通过血红素结合机制筛选具有潜在抗疟活性的分子的电化学方法。第一种方法适用于疏水环境,通过在 DMSO 溶液中使用溶液电化学,在碳电极上加入 Fe(III)-血红素和待测化合物。该方法提供了定义明确的伏安信号,这是血红素-配体(L)相互作用的特征。第二种方法涉及生物 pH 值下的水溶液和固定化颗粒的伏安法,通过将待测化合物的微粒膜浸入 Fe(III)-血红素溶液中,而无需预先孵育。这些方法学被应用于血红蛋白等大分子水平体系中的血红素结合活性的测试,或者更复杂的混合物,如全血、红细胞或溶血样品。