Binding of omeprazole to protein targets identified by monoclonal antibodies.
Binding of omeprazole to protein targets identified by monoclonal antibodies.
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DOI：
10.1371/journal.pone.0239464
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发表时间：
2020
期刊：
影响因子：
3.7
通讯作者：
Wang MM
中科院分区：
文献类型：
--
作者：
Cartee NMP;Wang MM
奥美拉唑是最常用的质子泵抑制剂（PPI），这类药物的治疗作用机制涉及在胃分泌细胞上的H+/K+ ATP酶泵中与半胱氨酸残基形成二硫键。奥美拉唑的唯一硫基与泵蛋白的选定半胱氨酸残基之间的共价键导致胃中酸分泌的抑制，这是一种改善胃食管反流和消化性溃疡疾病的作用。质子泵抑制剂，虽然适用于特定条件时，短暂使用，是与各种不良反应时，长期使用。这些潜在脱靶效应的机制仍不清楚。事实上，PPI可能与非典型靶蛋白（非泵分子）相互作用，导致意外的病理生理学效应，但很少有研究描述PPI的脱靶结合。在这里，我们描述了成功克隆单克隆抗体对蛋白结合奥美拉唑。我们开发并使用单克隆抗体来表征奥美拉唑的蛋白质靶范围、奥美拉唑结合蛋白的稳定性以及半胱氨酸参与奥美拉唑与靶点的结合。我们证明了奥美拉唑靶向广泛的不同蛋白质。蛋白质复合物，检测蛋白质印迹，耐热，洗涤剂和还原剂。奥美拉唑与不含半胱氨酸的蛋白质发生反应，不受半胱氨酸烷基化的完全抑制，在中性pH值下发生，并诱导蛋白质多聚化。至少有两种其他临床使用的PPI，雷贝拉唑和泰妥拉唑，能够以类似的方式与蛋白质结合。我们得出结论，奥美拉唑结合多种蛋白质，并能够形成高度稳定的复合物，不依赖于药物和蛋白质靶点之间的二硫键。通过这些抗体进行的进一步研究可能揭示PPI-蛋白复合物是否是长期使用PPI的脱靶不良反应的基础。
Omeprazole is the most commonly used proton pump inhibitor (PPI), a class of medications whose therapeutic mechanism of action involves formation of a disulfide linkage to cysteine residues in the H+/K+ ATPase pump on gastric secretory cells. Covalent linkage between the sole sulfur group of omeprazole and selected cysteine residues of the pump protein results in inhibition of acid secretion in the stomach, an effect that ameliorates gastroesophageal reflux and peptic ulcer disease. PPIs, though useful for specific conditions when used transiently, are associated with diverse untoward effects when used long term. The mechanisms underlying these potential off-target effects remain unclear. PPIs may, in fact, interact with non-canonical target proteins (non-pump molecules) resulting in unexpected pathophysiological effects, but few studies describe off-target PPI binding. Here, we describe successful cloning of monoclonal antibodies against protein-bound omeprazole. We developed and used monoclonal antibodies to characterize the protein target range of omeprazole, stability of omeprazole-bound proteins, and the involvement of cysteines in binding of omeprazole to targets. We demonstrate that a wide range of diverse proteins are targeted by omeprazole. Protein complexes, detected by Western blotting, are resistant to heat, detergents, and reducing agents. Reaction of omeprazole occurs with cysteine-free proteins, is not fully inhibited by cysteine alkylation, occurs at neutral pH, and induces protein multimerization. At least two other clinically used PPIs, rabeprazole and tenatoprazole, are capable of binding to proteins in a similar fashion. We conclude that omeprazole binds to multiple proteins and is capable of forming highly stable complexes that are not dependent on disulfide linkages between the drug and protein targets. Further studies made possible by these antibodies may shed light on whether PPI-protein complexes underlie off-target untoward effects of chronic PPI use.
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影响因子：
2.7
作者：
Schumock, Glen T.;Li, Edward C.;Vermeulen, Lee C.
通讯作者：
Vermeulen, Lee C.
影响因子：
29.4
作者：
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DOI：
10.1001/jama.2015.13766
发表时间：
2015-11-03
期刊：
JAMA
影响因子：
--
作者：
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通讯作者：
Giovannucci EL
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29
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影响因子：
3.7
作者：
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