Department of Chemistry, East Carolina University, Greenville, NC, United States.
Department of Chemistry, East Carolina University, Greenville, NC, United States.
Methods Enzymol. 2024;704:59-87. doi: 10.1016/bs.mie.2024.05.011. Epub 2024 Jun 18.
This Chapter describes methods for the biosynthetic substitution of the mononuclear, non-heme iron in plant and animal lipoxygenases (LOXs). Substitution of this iron center for a manganese ion results in an inactive, yet faithful structural surrogate of the LOX enzymes. This metal ion substitution permits structural and dynamical studies of enzyme-substrate complexes in solution and immobilized on lipid membrane surfaces. Representative procedures for two LOXs, soybean lipoxygenase (SLO) from plants and human epithelial 15-lipoxygenase-2 (15-LOX-2) from mammals, are described as examples.
本章介绍了植物和动物脂氧合酶（LOXs）中单核、非血红素铁的生物合成取代方法。用锰离子取代这个铁中心会得到 LOX 酶的无活性但忠实的结构替代物。这种金属离子取代允许在溶液中和固定在脂质膜表面上对酶-底物复合物进行结构和动力学研究。作为示例，描述了来自植物的大豆脂氧合酶（SLO）和来自哺乳动物的人上皮 15-脂氧合酶-2（15-LOX-2）这两种 LOX 的代表性程序。
