COBALT(II)-RADICAL PAIR DYNAMICS IN B12 ENZYME CATALYSIS

一种新兴的酶类，它能利用酶的极端活性 电子缺乏的自由基物种中进行最多的一些 最近发现了生物学中的疑难化学反应。 这些酶实现的区域和立体选择性是长期挑战的。 认为激进的反应是非特定的。这门课包括 以下是：核糖核苷酸还原酶，它催化第一个 DNA生物合成中独特的一步，前列腺素H合酶，靶标 阿司匹林和其他非类固醇抗炎药，以及 B12辅酶依赖酶家族，催化代谢产物 共价键重排。催化反应中的共同第一步 是金属配位或金属辅助生成缺电子的吗 有机自由基。这种引发剂自由基从 底物形成基于底物的自由基，开启新的反应 一种便于对产品进行重新排列的通道。一个 具有挑战性的问题是如何稳定自由基以防止重组 用金属制成的。与电荷分离反应有相似之处 在光合作用和呼吸生物能复合体中， 已经建立了基本的性能原则。然而， 与简单的电子隧穿分离不同 弱相互作用的氧化还原位置，金属-自由基分离涉及大- 重核的标度运动和相对较强的电子 互动。阐明蛋白质和蛋白质的基本原理 辅因子引导自由基稳定和随后的底物自由基 重新安排将是拟议研究的持续重点。这个 腺苷钴胺依赖系统和乙醇胺脱氨酶 具体地说，之所以被选中进行审查，是因为同源切割 钴-碳键形成CoII金属和5‘- 腺苷引发剂自由基可由可见激光脉冲触发， 允许自由基对状态的相干制备 催化条件。自由基对分离与底物自由基 重排将通过脉冲EPR的时间分辨技术进行跟踪 光谱学。动态电子自旋与电子核超精细 相互作用将被测量并用于构建详细的分子 机制。开发的洞察力和技术将促进 其他酶中瞬时自由基中间体的鉴定 反应，表明编程的特定部位的自由基的设计 体内反应，并协助治疗努力对抗 具有生物破坏性的自由基。

An emergent class of enzymes that harness the extreme reactivity of electron-deficient free radical species to carry out some of the most difficult chemical reactions in biology has been recently identified. The regio- and stereo-selectivity achieved by these enzymes defies long- held ideas that radical reactions are non-specific. This class includes the following: ribonucleotide reductases, which catalyze the first unique step in DNA biosynthesis, prostaglandin H-synthase, the target of aspirin and other non-steroidal anti-inflammatory drugs, and the family of B12 coenzyme-dependent enzymes, which catalyze metabolite covalent bond rearrangements. The common primary step in the catalysis is metallocenter- or metal-assisted generation of an electron-deficient organic radical. This initiator radical abstracts a hydrogen atom from the substrate to form a substrate-based radical, opening a new reaction channel that facilitates rearrangement to a product radical. A challenging issue is how the radical is stabilized against recombination with the metal. There are analogies with the charge separation reactions in photosynthetic and respiratory bioenergetic complexes, for which the rudimentary performance principles have been established. However, unlike simple electron-hole separation by electron tunneling among weakly interacting redox sites, metal-radical separations involve large- scale movement of heavy nuclei and relatively strong electronic interactions. Elucidating the basic principles of how protein and cofactors guide radical stabilization and ensuing substrate radical rearrangement will be sustained focuses of the proposed studies. The adenosylcobalamin-dependent systems, and ethanolamine deaminase specifically, have been selected for scrutiny because homolytic cleavage of the cobalt-carbon bond to form the CoII metalloradical and 5'- adenosyl initiator radical can be triggered by a visible laser pulse, allowing the coherent preparation of the radical pair state under catalytic conditions. The radical pair separation and substrate radical rearrangement will be tracked by time-resolved techniques of pulsed-EPR spectroscopy. Dynamic electron spin-spin and electron-nuclear hyperfine interactions will be measured and used to construct a detailed molecular mechanism. The insights and techniques developed will promote identification of transient radical intermediates in other enzyme reactions, indicate designs for programmed site-specific radicals reactions in vivo, and assist therapeutic efforts to combat biologically-destructive free radicals.