Department of Chemistry, Stanford University, Stanford, CA, United States; Stanford Synchrotron Radiation Lightsource, SLAC National Acceleration Laboratory, Stanford University, Menlo Park, CA, United States.
Department of Chemistry, Stanford University, Stanford, CA, United States.
Methods Enzymol. 2024;703:29-49. doi: 10.1016/bs.mie.2024.05.019. Epub 2024 Jun 21.
Non-heme iron enzymes play key roles in antibiotic, neurotransmitter, and natural product biosynthesis, DNA repair, hypoxia regulation, and disease states. These enzymes had been refractory to traditional bioinorganic spectroscopic methods. Thus, we developed variable-temperature variable-field magnetic circular dichroism (VTVH MCD) spectroscopy to experimentally define the excited and ground ligand field states of non-heme ferrous enzymes (Solomon et al., 1995). This method provides detailed geometric and electronic structure insight and thus enables a molecular level understanding of catalytic mechanisms. Application of this method across the five classes of non-heme ferrous enzymes has defined that a general mechanistic strategy is utilized where O activation is controlled to occur only in the presence of all cosubstrates.
非血红素铁酶在抗生素、神经递质和天然产物生物合成、DNA 修复、缺氧调节以及疾病状态中发挥着关键作用。这些酶一直难以用传统的生物无机光谱方法来研究。因此，我们开发了变温变场磁圆二色性（VTVH MCD）光谱法，以实验确定非血红素亚铁酶的激发和基态配体场状态（Solomon 等人，1995 年）。该方法提供了详细的几何和电子结构信息，从而使我们能够从分子水平上理解催化机制。该方法在五类非血红素亚铁酶中的应用，定义了一种普遍的机制策略，其中 O 激活仅在所有共底物存在的情况下才被控制发生。
