Identification of ligands from natural products as inhibitors of glutathione S‐transferases using enzyme immobilized mesoporous magnetic beads with high‐performance liquid chromatography plus quadrupole time‐of‐flight mass spectrometry and molecular docking

Abstract

Multidrug resistance is recognized as one of the main reasons leading to the failure of chemotherapy. Studies have shown that glutathione S‐transferase inhibitors could be regarded as multidrug resistance reversal agents. Herein, a method of applying enzyme immobilization, molecular docking, and high‐performance liquid chromatography coupled with quadrupole time‐of‐flight mass spectrometry was employed to screen glutathione S‐transferase inhibitors from natural products. Magnetic mesoporous silica microspheres were synthesized and modified with a poly(dopamine) layer, which has a large quantity of amino, enabling further non‐covalent binding with glutathione S‐transferase. Moreover, the immobilization conditions, namely, potential of hydrogen, catalase concentration, reaction temperature and reaction time, were optimized. In total, six potential compounds were isolated and identified from Perilla frutescens (L.) Britt leaves and green tea and molecular docking was applied to identify the binding site. Rosmarinic acid, (−)‐epigallocatechin‐3‐O‐gallate and (−)‐epicatechin‐3‐gallate showed higher binding affinity than the compounds, and their half maximal inhibitory concentration values were further determined. The results suggested that this proposed method was effective and convenient for identifying glutathione S‐transferase inhibitors from natural products.

Ligand fishing as a tool to screen natural products with anticancer potential

“…In this sense, IOMN silica-coating is an interesting procedure to enhance the chemical stability and reduce agglomeration and eventual cytotoxicity of this material [27]. In a sophisticated experiment, Yi et al [28] fabricated a polydopamine-functionalized silica-coated iron oxide mesoporous magnetic particle meant to immobilize glutathione-S-transferase (GST) on its surface. The authors used a hexadecyltrimethylammonium chloride template to create mesopores in the silica-coated IOMN, F I G U R E 5 Ligand fishing using magnetic particles (MPs).…”

Biomimetic mineralization synthesis of poly(sodium 4‐styrenesulfonate)‐mediated calcium carbonate magnetic microsphere for kallikrein immobilization

“…Immobilizations of enzymes are expected to enhance their reusability at a wide pH and temperature range [1, 2], which have been widely used in biosensors, food chemistry, and biofuel production [3]. There were several methods reported for enzymes’ immobilization, including physical methods such as entrapment [4] and noncovalent binding [5], as well as chemical methods such as covalent [6−10] and cross‐linking [11]. Nevertheless, the immobilization processes of enzymes are complicated in most cases.…”

“…Immobilizations of enzymes are expected to enhance their reusability at a wide pH and temperature range Abbreviations: AEBSF, 4-(2-aminoethyl)-benzenesulfonyl fluoride hydrochloride; BET, Brunauer-Emmett-Teller; EDX, energy dispersive X-ray spectroscopy; IC 50 , half-maximal inhibitory concentration; PSS, poly(sodium 4-styrenesulfonate); TGA, thermogravimetric analysis; VSM, vibrating sample magnetometry; XRD, X-ray diffraction [1,2], which have been widely used in biosensors, food chemistry, and biofuel production [3]. There were several methods reported for enzymes' immobilization, including physical methods such as entrapment [4] and noncovalent binding [5], as well as chemical methods such as covalent [6−10] and cross-linking [11]. Nevertheless, the immobilization processes of enzymes are complicated in most cases.…”

Development and evaluation of poly adenosine 5′‐diphosphate‐ribose polymerase 1 immobilization‐based receptor chromatography

“…We packed the gel into a stainless‐steel column, screened the bioactive component in YHD extract, and identified the compounds by HPLC‐MS/MS [22]. The binding mode of the bioactive compound and PARP1 was confirmed by molecular docking [23]. We expect the method can provide a universal way for drug discovery and shorten the time in lead compound screening.…”